Fig. 3: Shear-stress-induced autophagy and cell size regulation depend on PI3KC2α, but not on PI3KC3/VPS34. | Nature Communications

Fig. 3: Shear-stress-induced autophagy and cell size regulation depend on PI3KC2α, but not on PI3KC3/VPS34.

From: PI3KC2α-dependent and VPS34-independent generation of PI3P controls primary cilium-mediated autophagy in response to shear stress

Fig. 3

a, c Representative confocal images of polarized HK2 cells (siCTRL), compared to PI3KC2α knocked down cells (siPI3KC2α) and PI3KC3/VPS34 knocked down cells (siVPS34), upon static (ctrl, 24 h), shear stress (24 h) and starvation conditions (24 h, see Methods section for more details), immunostained for LC3, F-actin and DAPI. Scale bar, 10 μm. b, d Quantifications of (a) and (c). Bar graphs denote average number of LC3 puncta and average cell areas (mean ± SEM, N = 100 cells, from three independent experiments). NS: not significant, ***p < 0.001 in two-tailed Student’s t test. e, g Cortex zone of kidneys from wild type (WT) and PI3KC2+/− mice, immunostained for LC3, LTL (Lotus Tetragonolobus Lectin) and DAPI, and quantification of LC3 puncta (mean ± SEM, from three independent experiments). Scale bar, 10 μm. *p < 0.05 in two-tailed Student’s t test. f, h PAS (Periodic Acid Schiff) staining of the cortex zone of kidneys from wild type and PI3KC2+/− mice and quantification of tubular cell area (mean ± SEM, from 3 independent experiments). Scale bar, 50 μm. *p < 0.05 in two-tailed Student’s t test.

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