From 60535f8014acf7d64449b934b90d02e6c700336d Mon Sep 17 00:00:00 2001 From: Cynthia Condra Date: Sat, 5 Sep 2026 14:52:10 +0000 Subject: [PATCH 1/2] plotPCA: write and plot the per-sample loadings in the untransposed layout The re-implemented plot_pca set Wt = U * S for both layouts. With samples as the variables (the default, untransposed layout) U * S holds the PC scores of the rows of the matrix -- one row per selected bin -- so the table's "Component i" row and the plotted points were the first rows' scores across components, labelled with the sample names. The help documents "the loadings for each sample in each principal component"; those are the rows of V^T. --transpose (samples as observations, each sample's projection U * S) was already correct. The "not enough principal components to plot N samples" exit is removed: every component row has one loading per sample, so --ntop below the sample count plots fine as long as the requested PCs exist (the remaining check). The golden plotPCA PNGs/TSVs showed the bin scores and are regenerated. Test: test_plotPCA_default_table_holds_sample_loadings (rows of the default table are orthonormal and PC1 equals a numpy SVD of the same standardised rows); test_plotPCA_ntop_smaller_than_samples now expects one loading per sample (2 x 8 table) and test_plotPCA_requested_PC_beyond_ntop_errors_cleanly keeps the clean-exit coverage. Co-Authored-By: Claude Fable 5.1 Claude-Session: https://claude.ai/code/session_01TaHntBDKuZJpMAAMenkC44 --- CHANGES.txt | 1 + pydeeptools/deeptools/correlation.py | 20 ++++------ pydeeptools/deeptools/test/test_plotPCA.py | 36 +++++++++++++----- .../test_plotPCA/test_plotPCA_default.png | Bin 45174 -> 48393 bytes .../test_plotPCA/test_plotPCA_default.tsv | 12 +++--- .../test/test_plotPCA/test_plotPCA_ggplot.png | Bin 54586 -> 59789 bytes .../test/test_plotPCA/test_plotPCA_ggplot.tsv | 12 +++--- 7 files changed, 48 insertions(+), 33 deletions(-) diff --git a/CHANGES.txt b/CHANGES.txt index 56dfe7914d..552cf3488a 100755 --- a/CHANGES.txt +++ b/CHANGES.txt @@ -16,6 +16,7 @@ * alignmentSieve output order matches input order exactly * --missingDataAsZero no longer takes bases exceeding chromosome bounds as 0 values but rather purges the bins * large scale values precision slightly altered with new backend (f32 vs f64) +* plotPCA (untransposed, the default) again writes and plots the loadings of each sample on the principal components (rows of V^T); the re-implementation wrote the PC scores of the first rows of the matrix under the sample names. --transpose was correct 3.5.6 * minimal supported python version raised to 3.9 (numpy >= 2 support); NaN handling switched to np.nan diff --git a/pydeeptools/deeptools/correlation.py b/pydeeptools/deeptools/correlation.py index cd7a35acb6..cb225eb2bd 100644 --- a/pydeeptools/deeptools/correlation.py +++ b/pydeeptools/deeptools/correlation.py @@ -497,19 +497,22 @@ def plot_pca(self, plot_filename=None, PCs=[1, 2], plot_title='', image_format=N U *= signs Vt *= signs[:, None] - # Projected coordinates: U * S == X @ V - Wt = U * S - # Eigenvalues and % variance explained. eigenvalues = (S ** 2) / (n_samples - 1) variance = eigenvalues / eigenvalues.sum() pvar = variance / variance.sum() if self.transpose: - # With samples as observations, U * S already gives each sample's + # Samples are the observations: U * S == X @ V gives each sample's # projection onto the PCs (rows=samples, cols=components). Orient as # (components, samples) to match the indexing used below. - Wt = Wt.T + Wt = (U * S).T + else: + # Samples are the variables: what is written and plotted per sample + # is its loading on each component, i.e. the rows of V^T + # (components, samples). U * S would be the scores of the rows + # (bins) of the matrix, one per selected bin, not per sample. + Wt = Vt if plot_filename is not None: n = n_bars = len(self.labels) @@ -519,13 +522,6 @@ def plot_pca(self, plot_filename=None, PCs=[1, 2], plot_title='', image_format=N if max(PCs) > eigenvalues.size: sys.exit("Cannot plot PC{}: only {} principal component(s) are " "available. Reduce --PCs or increase --ntop.\n".format(max(PCs), eigenvalues.size)) - # In the untransposed layout each point is a sample indexed along - # the component axis, so there must be at least as many components - # as samples (i.e. enough usable rows / a large enough --ntop). - if not self.transpose and Wt.shape[1] < n: - sys.exit("Not enough principal components ({}) to plot {} " - "samples; increase --ntop to at least the sample " - "count.\n".format(Wt.shape[1], n)) markers = itertools.cycle(matplotlib.markers.MarkerStyle.filled_markers) if cols is not None: colors = itertools.cycle(cols) diff --git a/pydeeptools/deeptools/test/test_plotPCA.py b/pydeeptools/deeptools/test/test_plotPCA.py index a284677764..dac41372ae 100644 --- a/pydeeptools/deeptools/test/test_plotPCA.py +++ b/pydeeptools/deeptools/test/test_plotPCA.py @@ -158,22 +158,21 @@ def test_plotPCA_ntop_zero_uses_all_rows(): def test_plotPCA_ntop_smaller_than_samples(): """When --ntop is below the sample count the table is truncated to the - number of retained components (rows).""" - # plot=False: the numeric table is well-defined even with 2 features. - data = _run_pca(["--ntop", "2"], plot=False) - assert data.shape == (2, 4) + number of retained components (rows); every row still holds one loading + per sample, so the plot of PC1 vs PC2 is well-defined.""" + data = _run_pca(["--ntop", "2"]) + assert data.shape == (2, 8) np.testing.assert_array_equal(data[:, 0], np.arange(1, 3)) # First component carries all the variance for the 2-feature case. np.testing.assert_allclose(data[0, -1], 12.0, rtol=1e-6) assert abs(data[1, -1]) < 1e-6 -def test_plotPCA_ntop_below_samples_plot_errors_cleanly(): - """Plotting with fewer retained components than samples cannot lay out the - scatter; the tool must exit with a clear message rather than crash with an - IndexError (previously a bug at correlation.py's scatter loop).""" +def test_plotPCA_requested_PC_beyond_ntop_errors_cleanly(): + """Asking for a component that the retained rows cannot provide must exit + with a clear message rather than crash with an IndexError.""" plotfile = NamedTemporaryFile(suffix='.png', prefix='deeptools_testfile_', delete=False) - args = "-in {0}test_samples.npz -o {1} --ntop 2".format(TEST_DATA, plotfile.name).split() + args = "-in {0}test_samples.npz -o {1} --ntop 2 --PCs 1 3".format(TEST_DATA, plotfile.name).split() try: with pytest.raises(SystemExit) as exc: deeptools.plotPCA.main(args) @@ -183,6 +182,25 @@ def test_plotPCA_ntop_below_samples_plot_errors_cleanly(): os.remove(plotfile.name) +def test_plotPCA_default_table_holds_sample_loadings(): + """In the default (untransposed) layout the table row for component i is + the loading of every sample on PC i, i.e. row i of V^T from the SVD of the + standardised (ntop rows x samples) matrix: the rows are orthonormal and + equal, up to the per-component sign, to a numpy PCA on the same rows. + (The bin scores U*S, one row per selected bin, are neither.)""" + data = _run_pca() + W = data[:, 1:7] + np.testing.assert_allclose(W @ W.T, np.eye(6), atol=1e-6) + + mat = np.load(TEST_DATA + "test_samples.npz")["matrix"].astype(float) + m = mat[np.argpartition(mat.var(axis=1), -500)[-500:]] + m = (m - m.mean(axis=0)) / m.std(axis=0) + _, _, Vt = np.linalg.svd(m - m.mean(axis=0), full_matrices=False) + # PC1 dominates and is well separated, so its loadings are stable + # across BLAS backends; compare it sign-invariantly. + np.testing.assert_allclose(_sign_fix(W[:1].T)[:, 0], _sign_fix(Vt[:1].T)[:, 0], rtol=1e-5, atol=1e-8) + + def test_plotPCA_PCs_selection_does_not_change_table(): """--PCs only selects which components are drawn; the numeric table always contains every component, so it is independent of --PCs.""" diff --git a/pydeeptools/deeptools/test/test_plotPCA/test_plotPCA_default.png b/pydeeptools/deeptools/test/test_plotPCA/test_plotPCA_default.png index e27190e1c44714e8ca1a3b87a487d5014fe94357..deb6ececb14ab19b5295de6806b2ec99664c5f90 100644 GIT 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z&Bq#-d-OLTy;ODIYYi?4Kks$=a)S;OVtEg?fH-@L&j}6}tghB?VXs2_Z`WL#M3N}K zo4q#x(ULn(I$z#3A^I&;e>^Uhuuig}4|(d;+iyt6Wptz9!=qO>NJ9PI$}^$^oXX4i z0h6_W*I?|o@qJ^dpe+1^S~G7lYabxL&OjkmxHZ`52@mVL`>KTm$Q)fvexyGv@wYTb z5Gd-iEPtKo%_GWyHz)tO2wC@hrsf4p5Fr5D`ZcoTGeKpirnNu!8xb+cQt z(Y%Ph^!2D@Ij59$Fw?9^$wKQ|!j#3-!Q(+9l!o>9l(w!9sq3)M9(uLqnibzFnDiZu zA)_%GA5u;gYf6vJ`$%>CWr#oDktMSlCtVZQ$6pMCGH=Ue2Hr}Vq0TVyI-?U!=fY}LkL7u&K+p0cj~4lHuu%m+J>&;x0A;NI|F4fZn~VjO!*gdl SDRIa#C50RRrvG#6@qYoY!riX` diff --git a/pydeeptools/deeptools/test/test_plotPCA/test_plotPCA_ggplot.tsv b/pydeeptools/deeptools/test/test_plotPCA/test_plotPCA_ggplot.tsv index 9822010ab4..b568eac027 100644 --- a/pydeeptools/deeptools/test/test_plotPCA/test_plotPCA_ggplot.tsv +++ b/pydeeptools/deeptools/test/test_plotPCA/test_plotPCA_ggplot.tsv @@ -1,7 +1,7 @@ Component wt1 wt2 wt3 kd1 kd2 kd3 Eigenvalue -1 -1.9593953629718677 0.05992632052266172 0.015514438980501043 0.360872626954001 -0.06417725736657884 0.012713770441428606 5.807692278755935 -2 -3.3594106951252853 -0.1453930968029286 -0.134748642812576 -0.012411165664076072 -0.12363598259528208 -0.060663076611996596 0.07423028883557839 -3 -2.335822964750958 0.09987722598713754 -0.03413541974064124 -0.07165342296216681 0.05170385813600347 0.1373257036829112 0.0489717777349372 -4 -0.30789783922401226 0.004185214180292798 -0.013424388619808516 0.051462455776524196 -0.07942877370987256 0.0745864450320923 0.036809415525390125 -5 -1.2801086861541353 -0.07795301634249786 -0.04768720563614555 -0.02321427427821972 -0.032405677000559435 0.07420665888684398 0.026706723301448902 -6 -3.127415384537607 0.17202014078313335 0.08493798982333509 0.01429353530978131 0.018508562924871238 0.15753396743432724 0.017613563942900885 +1 0.41015948328129415 0.40714083168340265 0.4051015001575071 0.4088383560914448 0.40895832745985133 0.4092708162212304 5.807692278755935 +2 -0.10564992452808587 -0.13903863717051682 -0.7036293245034178 -0.031273785139079135 0.5057293580802189 0.46655320303121317 0.07423028883557847 +3 -0.3072826224468924 0.8073633171644164 -0.366048315157121 0.2231968254778915 -0.12059197662562816 -0.23535345181275683 0.048971777734937216 +4 -0.16638072203492837 -0.3999296950298342 -0.0911203187723264 0.8821546210006602 -0.10280300047534902 -0.12371559972677873 0.03680941552539014 +5 0.8255039368543798 0.008574695725392222 -0.4455301735837803 0.057473412505828825 -0.3107944624876728 -0.1416905014440879 0.026706723301448895 +6 -0.13055798419418163 0.05524299298321737 0.0026704255973279928 0.023587232780211485 -0.6747494521159683 0.7239147136476167 0.017613563942900895 From 5df8588a1d4c4abb653a544b6a35b0cfc85dd0bb Mon Sep 17 00:00:00 2001 From: Claude Date: Sat, 5 Sep 2026 16:43:18 +0000 Subject: [PATCH 2/2] plotPCA Galaxy wrapper test: expect sample loadings, not bin scores The wrapper's second test pinned the values plotPCA wrote before this change (1.99949, 0.96079: PC scores of the first bins). With the two identical input samples the loadings are +-1/sqrt(2) on PC1 with eigenvalue 3 and a degenerate PC2 (BLAS-dependent noise), so the assertions now check the PC1 loadings sign-agnostically and only the structure of the second row. The golden PNGs and tabular are regenerated from the corrected output (the PNGs compare by sim_size). Co-Authored-By: Claude Fable 5.1 Claude-Session: https://claude.ai/code/session_01TaHntBDKuZJpMAAMenkC44 --- galaxy/wrapper/plotPCA.xml | 15 ++++++++------- galaxy/wrapper/test-data/plotPCA_result1.png | Bin 48558 -> 47747 bytes galaxy/wrapper/test-data/plotPCA_result2.png | Bin 52610 -> 47747 bytes .../wrapper/test-data/plotPCA_result2.tabular | 4 ++-- 4 files changed, 10 insertions(+), 9 deletions(-) diff --git a/galaxy/wrapper/plotPCA.xml b/galaxy/wrapper/plotPCA.xml index f5fe600e88..3f62bf9da6 100644 --- a/galaxy/wrapper/plotPCA.xml +++ b/galaxy/wrapper/plotPCA.xml @@ -79,16 +79,17 @@ - + - - + + diff --git a/galaxy/wrapper/test-data/plotPCA_result1.png b/galaxy/wrapper/test-data/plotPCA_result1.png index b043d20f58b55f2c8636cd13cea73fb9897142f5..9909578eeb0ca0d3deb2784fd90b77587f15060f 100644 GIT binary patch literal 47747 zcmeFa2T+x1*DZ+JhBme>W=t(8K@bH*5X7tpK@rIr5fBhif=I@I+A5$&GJ*+EGLkc> 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