Fig. 2: Cilia disassembly stimuli activate PI3K/AKT, MAPK, PKC and PKA signalling. | Nature Communications

Fig. 2: Cilia disassembly stimuli activate PI3K/AKT, MAPK, PKC and PKA signalling.

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

Fig. 2

a Layout of experimental induction and analysis of serum starvation-induced cilia assembly and stimulation-induced cilia disassembly. b Experimental design and workflow of phosphoproteomic experiment in hTERT-RPE1 cells. Cells were serum-starved for 48 h and stimulated with 10% serum or 2 μM LPA ± 0.5 μM GDC-0941 for 15 min or 2 h and processed for phosphoproteomic analysis. 10067 phosphosites from 2937 proteins were analysed by MSstats, 427 phosphosites were significantly regulated which were used for comparison with cilia proteome databases and for KSEA (n = 5 independent experiments). c Venn diagram showing phosphosites regulated by serum or 2 μM LPA ± 0.5 μM GDC-0941 of 48 h serum-starved hTERT-RPE1 cells relative to DMSO. 10067 phosphosites were quantified by phosphoproteomics of which 427 were differentially regulated. Phosphosites in cilia-associated proteins (as defined by SYSCILIA version 2 or CiliaCarta) are listed. d Heatmap displaying phosphosites from cilia-associated proteins (as defined by SYSCILIA) regulated by 15 min or 2 h serum or 2  μM LPA stimulation ± 0.5 μM GDC-0941 in 48 h serum-starved hTERT-RPE1 cells relative to DMSO. e Heatmaps displaying KSEA (using the OmniPath, Edges and PhosphoSitePlus database) of kinases for which the substrate groups were differentially regulated by 15 min or 2 h serum or 2 μM LPA stimulation ± 0.5 μM GDC-0941 in 48 h serum-starved hTERT-RPE1 cells relative to DMSO. Kinases for which the adjusted p-values (using the Kolmogorov–Smirnov test, adjusted for multiple comparisons with Benjamini-Hochberg principle (5% FDR)) relative to DMSO control were less than p = 0.05 were considered significantly regulated.

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