Fig. 3: Inhibition of PI3Kα or PI3Kβ impairs cilia disassembly in hTERT-RPE1 cells. | Nature Communications

Fig. 3: Inhibition of PI3Kα or PI3Kβ impairs cilia disassembly in hTERT-RPE1 cells.

From: A class I PI3K signalling network regulates primary cilia disassembly in normal physiology and disease

Fig. 3

a Inhibitors used. b Cells were serum-starved for 48 h, pre-treated with inhibitors or DMSO for 1 h and stimulated with serum in the presence of inhibitors for 24 h. % ciliated cells was scored, bars: mean ± SD, 100 cells/condition for n = 3,4 independent experiments, *p < 0.05, ****p < 0.0001 (one-way ANOVA, p = 3.3 × 10−7). c Cells were serum-starved for 48 h, pre-treated with inhibitors or DMSO for 1 h and then stimulated with LPA or serum in the presence of inhibitors for 2 h. Lysates were immunoblotted with pAKT(S473), AKT and GAPDH antibodies, representative of n = 3 independent experiments. d Cells were serum-starved for 48 h, pre-treated with inhibitors or DMSO for 1 h and then stimulated with serum in the presence of inhibitors for 2 h. Cells were stained with ARL13B and pS6RP(S240/244) antibodies and DAPI and imaged by confocal microscopy, arrows: cilia, bar: 10 μm, cells with low pS6RP(S240/244) are indicated by a white outline. pS6RP(S240/244) MFI was measured in ciliated cells and presented as a histogram. n = 223–254 cells/condition from 3 independent experiments ****p < 0.0001 (Kruskal-Wallis test, p = 2.59 × 10−53). e Cells were serum-starved for 48 h, pre-treated with inhibitors or DMSO for 1 h and then stimulated with LPA in the presence of inhibitors for 24 h. % ciliated cells was scored, bars indicate mean ± SD, 100 cells/condition for n = 3 independent experiment, *p < 0.05, **p < 0.01, ****p < 0.0001 (one-way ANOVA, p = 5.032 × 10−6). f Cells were serum-starved for 48 h, pre-treated with GSK2636771 or DMSO for 1 h and then stimulated with serum (left) or LPA (right) in the presence of inhibitors for 24 h. % ciliated cells was scored, bars: mean ± SD, 100 cells scored/condition for n = 6 (left), n = 5 (right) independent experiments, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 (one-way ANOVA, left 1.06 × 10−6, right p = 0.0043). g Pik3ca+/+/Pik3ca-/- MEFs were serum-starved for 48 h, ± 24 or 48 h serum stimulation. % ciliated cells was scored, bars: mean ± SD, 100 cells/condition for n = 3 independent experiments, *p < 0.05, **p < 0.01, ****p < 0.0001 (two-way ANOVA, left interaction p = 0.0155, row p = 2.072 × 10−6, column p = 0.2335, right interaction p = 0.00267, row p = 1.022 × 10−5, column p = 0.00598). Source data are provided as a Source Data file.

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