RPS6 Antibody - #AF6354

Fig. 5. Disordered iron homeostasis and inhibited mTORC1 activity upon exposure to CdCl2 and PCB77 at low dose. (A) The relative fluorescence intensity of CAeAM for measuring LIP to reflect intracellular iron availability (n = 3–4), and (B) Representative blots of FTH1 protein content to reflect iron storage. Analyses were performed after single or combined exposure to CdCl2 and PCB77 at 1 μM for 48 h. (C) Phosphorylated S6 and total S6 content to re- flect mTORC1 activity as measured by Western blot. Ratio of FTH1 to eIF2αP and ratio of pS6 to S6 in the control group were defined as 1. Analyses were performed after single or combined exposure to CdCl2 and PCB77 at 1 μM for 48 h. a- significantly different from the control group. Data were presented in mean ± SE. P < 0.05 was considered statistically significant.

FIGURE 6 Bergenin promotes fibroblasts autophagy mainly via inhibiting mTOR signaling pathway. Mlg and NIH-3 T3 cells were co-treated with bergenin (30 and 60 μM) and TGF-β1 (5 ngml1 ) for 24 h. (a,b) The expression of p-mTOR, mTOR, p-ULK1, ULK1, p-S6, and S6 in Mlg cells (a) and NIH-3 T3 cells (b). GAPDH was used as the internal control. Data are presented as the means ± SD. Experiments were performed in triplicate (n = 3). *p < .05; **p < .01; ***p < .001

FIGURE 5 Western blotting of downstream regulated proteins of CUDC-907 and immunohistochemistry staining. (A) The expression of c-Myc is regulated by the HDAC and PI3K/AKT/mTOR pathway, and both can be inhibited by CUDC-907. (B) Expression levels of proteins [c-Myc, p-AKT, p-S6, p-4EBP1, and acetyl histone H3K9 (H3K9ac)] in HCC cell lines affected by CUDC-907 were detected by western blotting. GAPDH was set as the internal reference. (C) Expression levels of proteins (same as figure 5B) from xenografted tumor with or without CUDC-907 treatment were detected. (D) Immunohistochemistry staining. Representative images of Ki67, c-Myc, and TUNEL staining of xenografts treated with CUDC-907 or vehicle. (E) Quantification of immunohistochemistry staining was calculated by the immunohistochemistry score method. The percentage of apoptotic area in tumor slides of TUNEL staining was calculated by IMAGE J software. *p < 0.05, **p < 0.001 (result compared with Vehicle group).

Fig. 5 Ketogenic diet activated mTORC2 in the heart of spontaneously hypertensive rats. a The phosphorylation of AMPK and the ratio of p-S6/S6 and p-Akt/Akt in the heart of Wistar rats and SHRs were measured with Western blots. The quantitative result were shown in b–d. Values are the mean ± SEM. n = 4 for each group. *P < 0.05, **P < 0.01

Fig. 5 H2O2 induces GC autophagy through the mTOR and Ras-cAMPPKA signaling pathway. Primary cultured GC were treated with H2O2 (200 μmol/l) for different time periods. Then, the GC samples were analyzed by immunoblotting using the corresponding antibody. a The expression of mTOR protein was determined by western blotting. b The expression of PI3K, AKT, and RPS6 was determined by western blotting.c The expression of Ras, cAMP, and PKA was determined by western blotting. d–j The relative expression of p-mTOR, p-PI3K, p-AKT, pRPS6, Ras, cAMP, and PKA were quantified by software Image J. Data represent as the means ± SE from three independent experiments. * p < 0.05, ** p < 0.01