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. 2015 Nov 19;2(12):1944–1956. doi: 10.1016/j.ebiom.2015.11.031

Fig. 3.

Fig. 3

p38 targets distinct phosphorylation sites of TSC2 in different CRC cell lines. A.WT, TSC1−/− or TSC2−/− MEF cells were treated with 10 μM SB202190 for 2 h and analyzed for P-S6K1 and P-S6 by immunoblot. B. CRC cells were transfected with two different TSC1 or TSC2 siRNAs, and then treated with 10 μM SB202190 for 2 h. TSC1, TSC2 and P-S6K1(T389) were analyzed by immunoblot. C. CRC cells were transfected with two different TSC1 or TSC2 siRNAs, and then treated with 10 μM SB202190 for 48 h. Cell growth was measured by SRB assay and expressed as percentage of SB202190/control (DMSO). Data represent means ± s.d. from three independent triplicate experiments. *P < 0.01 versus NC. D. Drug-sensitive CACO2, SW480 and RKO cells, and drug-resistant HCT116, SW620 and SW1116 cells were treated with 10 μM SB202190 for 2 h and analyzed for phosphorylation of TSC2 at S664, S1254, T1462 and S1798 by immunoblot. Boxed area highlights the different effects of SB202190 on TSC2 phosphorylation in drug-sensitive versus -resistant CRC cells. SE, short exposure, LE, long exposure. E. HCT116 and SW620 cells were treated with 10 μM SB202190 with or without Mek inhibitors 30 μM PD98059 or 10 μM U0126 for 2 h, and then analyzed for phosphorylation of TSC2, ERK, RSK, S6K1 and S6 by immunoblot.