Fig. 7: PDK1, PKCι, CEP170 and KIF2A drive cilia loss induced by PI3Kα activation. | Nature Communications

Fig. 7: PDK1, PKCι, CEP170 and KIF2A drive cilia loss induced by PI3Kα activation.

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

Fig. 7

hTERT-RPE1(Vector/PIK3CAH1047R) cells were serum-starved for 48 h with doxycycline and treated ± (a) GSK2334470, (b) MK2206 or (c) 229 for the final 24 h, or transfected with non-targeted control, (d) siPRKCI, (e) siCEP170 or (f) siKIF2A, fixed, stained with ARL13B and pericentrin antibodies and DAPI and the % ciliated cells scored, bars: mean ± SD, 100 cells scored/condition for n = 3 independent experiments, *p < 0.05, **p < 0.01, ***p < 0.001 ****p < 0.0001 (two-way ANOVA (a) interaction p = 0.0002, row factor p = 0.0002, column factor p = 0.0003, (b) interaction p = 0.0336, row factor p = 0.0006, column factor p = 0.0092, (c) interaction p = 0.002, row factor p = 1.196 × 10−5, column factor p = 7.744 × 10−4, (d) interaction p = 0.0018, row factor 4.962 × 10−5, column factor p = 0.4681, (e) interaction p = 0.0295, row factor p = 5.185 × 10−5, column factor p = 0.0295, (f) interaction p = 0.0403, row factor p = 0.0002, column factor p = 0.0264). g hTERT-RPE1(Vector/PIK3CAH1047R) cells were serum-starved for 48 h with doxycycline and treated with BYL719, TGX-221, GDC-0941, GSK2334470, MK2206, 229 or DMSO. Cells were fixed, stained with ARL13B and pericentrin antibodies and DAPI and the % ciliated cells scored, bars: mean ± SD, 100 cells scored/condition for n = 3 independent experiments, ***p < 0.001, ****p < 0.0001 (one-way ANOVA, p = 3.217 × 10−7). h hTERT-RPE1 cells were serum-starved for 48 h, pre-treated with BYL719, TGX-221, GDC-0941, GSK2334470, MK2206, 229 or DMSO for 1 h and then stimulated with serum in the presence of inhibitors for 24 h. Left, cells were fixed, stained with ARL13B and pericentrin antibodies and DAPI and the % ciliated cells scored, bars: mean ± SD, 100 cells scored/condition for n = 5 independent experiments, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 (one-way ANOVA, p = 2.092 × 10−9). Right, cells were lysed and immunoblotted with pAKT(S473), AKT and GAPDH antibodies, blots representative of n = 3 independent experiments. i Model for PI3K/PIP3-driven regulation of cilia disassembly. Source data are provided as a Source Data file.

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