Figure S1.
Disruption of focal adhesions does not lead to autophagy defects. (A–C) Migfilin depletion inhibits autophagy activity. (A) Immunoblotting analysis of p62 and LC3 protein levels in control (siCtrl) and migfilin knockdown (siMig #1 and siMig #2) KP4 cells under nutrient-rich (control), EBSS starvation (Starv.) condition. (B and C) Quantification analysis of p62 (B) and LC3-II (C) levels in A. Data represent mean ± SE, *P < 0.05, **P <0.01, ***P < 0.001, one-way ANOVA. n = 4 independent experiments. (D–G) Disruption of focal adhesions does not lead to autophagy defects. (D) Representative images of immunofluorescence staining for paxillin and Kindlin-2 in control (siCtrl) and paxillin knockdown (siPXN #1 and siPXN #2) KP4 cells. Scale bar: 10 µm. Quantification analysis was shown in the right panel. At least 22 cells in each group were analyzed. Data represent mean ± SE, ***P < 0.001, one-way ANOVA. n = 3 independent experiments. (E) Immunoblotting analysis of p62, LC3 and migfilin levels in control (siCtrl) and paxillin knockdown (siPXN #1 and siPXN #2) KP4 cells. Quantification analysis of p62, LC3-II and migfilin levels was shown in the right panel. Data represent mean ± SE, n.s., no significance, one-way ANOVA. n = 3 independent experiments. (F) Immunoblotting analysis of p62, LC3 and migfilin levels in control (siCtrl) and vinculin knockdown (siVin #1 and siVin #2) KP4 cells. Quantification analysis of p62, LC3-II and migfilin levels was shown in the right panel. Data represent mean ± SE, n.s., no significance, one-way ANOVA. n = 3 independent experiments. (G) KP4 cells were stably transfected with GFP-LC3 (green) under control condition and imaged by fluorescence microscopy. The number of GFP-LC3 puncta was measured. Representative images were shown in the left panel and quantification analysis was shown in the right panel. Scale bar: 10 µm (Magnify: 2 µm). At least 30 cells in each group were analyzed. Data represent mean ± SE, n.s., no significance, one-way ANOVA. n = 3 independent experiments. Source data are available for this figure: SourceData FS1.

Disruption of focal adhesions does not lead to autophagy defects. (A–C) Migfilin depletion inhibits autophagy activity. (A) Immunoblotting analysis of p62 and LC3 protein levels in control (siCtrl) and migfilin knockdown (siMig #1 and siMig #2) KP4 cells under nutrient-rich (control), EBSS starvation (Starv.) condition. (B and C) Quantification analysis of p62 (B) and LC3-II (C) levels in A. Data represent mean ± SE, *P < 0.05, **P <0.01, ***P < 0.001, one-way ANOVA. n = 4 independent experiments. (D–G) Disruption of focal adhesions does not lead to autophagy defects. (D) Representative images of immunofluorescence staining for paxillin and Kindlin-2 in control (siCtrl) and paxillin knockdown (siPXN #1 and siPXN #2) KP4 cells. Scale bar: 10 µm. Quantification analysis was shown in the right panel. At least 22 cells in each group were analyzed. Data represent mean ± SE, ***P < 0.001, one-way ANOVA. n = 3 independent experiments. (E) Immunoblotting analysis of p62, LC3 and migfilin levels in control (siCtrl) and paxillin knockdown (siPXN #1 and siPXN #2) KP4 cells. Quantification analysis of p62, LC3-II and migfilin levels was shown in the right panel. Data represent mean ± SE, n.s., no significance, one-way ANOVA. n = 3 independent experiments. (F) Immunoblotting analysis of p62, LC3 and migfilin levels in control (siCtrl) and vinculin knockdown (siVin #1 and siVin #2) KP4 cells. Quantification analysis of p62, LC3-II and migfilin levels was shown in the right panel. Data represent mean ± SE, n.s., no significance, one-way ANOVA. n = 3 independent experiments. (G) KP4 cells were stably transfected with GFP-LC3 (green) under control condition and imaged by fluorescence microscopy. The number of GFP-LC3 puncta was measured. Representative images were shown in the left panel and quantification analysis was shown in the right panel. Scale bar: 10 µm (Magnify: 2 µm). At least 30 cells in each group were analyzed. Data represent mean ± SE, n.s., no significance, one-way ANOVA. n = 3 independent experiments. Source data are available for this figure: SourceData FS1.

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