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. 2023 Jul 24;13:11922. doi: 10.1038/s41598-023-39093-7

Figure 5.

Figure 5

The knockdown of GGCT down-regulates pSTAT3 and c-Met and activates the RB protein via AMPK. (a) A Western blot analysis of GGCT, c-Met, pAMPK Thr172, pSTAT3 Ser727, and their non-phosphorylated forms in PC3 cells treated with control siRNA, GGCT siRNA#2, and/or the indicated AMPK siRNAs for 72 h. Vinculin is shown as a loading control. All full-length blots are presented in Supplementary Fig. S8. (b) The expression of human-MET mRNA, as assessed by qRT-PCR, in PC3 cells at 72 h post-transfection with control siRNA, GGCT siRNA#2, and/or AMPKα siRNA#2. A two-tailed Student’s t-test was used (N = 3 per group, *p < 0.05). The error bars represent the S. D. (c) A Western blot analysis of pMEK Ser217/221, pERK Thr202/Tyr204, pRB Ser780, and their non-phosphorylated forms in PC3 cells treated with control siRNA, GGCT siRNA#2, and/or the indicated AMPK siRNAs for 72 h. Vinculin is shown as a loading control. All full-length blots are presented in Supplementary Fig. S8. (d) The viable cell numbers of PC3 cells treated with control siRNA, GGCT siRNA#2, and/or the indicated AMPK siRNAs for 72 h were analyzed by the trypan blue dye exclusion test. A one-way ANOVA with Dunnett’s test was used (N = 3 per group, *p < 0.05). The error bars represent the S. D. (e) Fractions in the S phase of PC3 cells at 48 h post-transfection with indicated siRNAs were measured by a BrdU incorporation assay. A one-way ANOVA with Dunnett’s test was used (N = 3 per group, **p < 0.01, ***p < 0.001). The error bars represent the S. D.