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. 2016 May 18;7(27):41445–41459. doi: 10.18632/oncotarget.9441

Figure 4. Inhibition of ROCK repressed the activity of JAK2 and STAT3.

Figure 4

(A) Knockdown of RhoE by siRNA induced phosphorylation of both JAK2 and STAT3 in BEL-7402 and SMMC-7721 cells. (B) Treatment with Y27632 for 24 hours inhibited the phosphorylation of JAK2 and STAT3 in BEL-7402 and SMMC-7721 cells. (C) Schematic diagram showing the construct which expressed firefly luciferase under the control of STAT3-binding elements. Both Y27632 and ROCK2 knockdown by siRNA reduced the luciferase activity in BEL-7402 cells. SV40-driven expression of Renilla luciferase was used as normalization control. (D) Treatment with Y27632 for 24 hours decreased the protein level of c-myc, a transcriptional target of STAT3, in BEL-7402 and SMMC-7721 cells. (E) Representative IHC staining of p-STAT3 (Y705) in xenograft tumors harvested from subcutaneous tumor injection model (Figure 3B). Y27632 remarkably diminished the p-STAT3 (Y705) signals; the p-STAT3 stain was nearly totally abolished in the combined Y27632 and cisplatin treatment group. Original magnification, ×20; scale bars: 200 μm. 10 μM Y27632 was used. In all panels, 3 experiments are represented. P-value was calculated using Student's t test.