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. 2003 Oct 15;112(8):1223–1233. doi: 10.1172/JCI17222

Figure 5.

Figure 5

Reduced PI3K-Akt signaling in Tsc2–/–TP53–/– and Tsc2–/– cells. (a) Immunoblot analysis of cell extracts showing reduced pAkt in Tsc2–/–TP53–/– in comparison to control TP53–/– cells at all time points after 10% serum addition. pAkt S473, pAkt (Ser473). (b) Immunoblot analysis showing reduced pAKT in serum-starved Tsc1–/– and Tsc2–/–TP53–/– cells with and without treatment with calyculin A. (c) Autoradiography demonstrates reduced PI3K activity in Tsc2–/–TP53–/– cells in comparison with TP53–/– cells, in response to PDGF. p-tyrosine, phosphotyrosine; PIP, phosphoinositide phosphate. (d) Ruffling activity, indicated by arrowheads, in response to PDGF was reduced in Tsc2–/–TP53–/– cells in comparison to control TP53–/– cells or a revertant TSC2-expressing cell line. Error bars (n = 3) depict the SD. (e) YPH-Akt translocation is reduced in Tsc2–/–TP53–/– cells in comparison to control TP53–/– cells or a revertant TSC2-expressing cell line. PDGF stimulation leads to uniform YFP staining of the plasma membrane in Tsc2+/+ and revertant cells, but not in Tsc2–/– cells. The staining intensity in cross-sections of the same cells is shown in the graphs at right. AU, arbitrary units.