Fig 2: Effects of stress-mimicking cortisol concentrations on Rack1 and mBDNF expression. Analysis of Rack1 mRNA and protein levels (A-D) in SH-SY5Y cells exposed to either 1 or 5 µM cortisol or vehicle control (DMSO < 0.1%) for 24 h (A-B) and 48 h (C-D). Analysis of mBDNF protein levels (E-F) in SH-SY5Y cells treated with either 1 or 5 µM cortisol or vehicle control (DMSO < 0.1%) for 24 h (E) and 48 h (F). Data are expressed as percentage of CTRL (set at 100%). *p<0.05, **p < 0.01 vs CTRL (one-way ANOVA with Tukey’s multiple comparison test).

Fig 3: VRB modulates mTOR/4E-BP1 to promote eIF4F complex remodeling and SG formationA. VRB disrupts eIF4F complex formation and enhances eIF4E/4E-BP1 interactions. U2OS cells without (−) or with VRB (150 μM), SA (100 μM), DOX (150 μM) or hydrogen peroxide (H2O2, 1mM) treatment (1hour) were lysed and subjected to m7GTP-sepaharose pull down to isolate cap-bound complexes. Both the input (input) and precipitated (m7GTP) fractions were subjected to western blotting and probed against eIF4G, eIF4E and non-phosphorylated 4E-BP1 (non-ph-4EBP1). B. The same as in 5A, except that U2OS cells were treated with indicated VAs and levels of non-phosphorylated 4E-BP1 (non-ph-4EBP1) were compared to control treatment (−). Levels of eIF4G and eIF4E were served as controls. C. VRB treatment does not affect ph-RPS6 status. Whole cell lysates from VRB-, SA-, DMSO-treated and untreated (−) U2OS cells were subjected to western blotting using ph-eIF2α-, ph-RPS6 and non-ph-4E-BP1-specific antibodies. RACK1 was used as loading control. D. VRB-induced SG formation in MEFs carrying non-phosphorylatable variant of RPS6 (ph-RPS6−/−). WT and ph-RPS6−/− MEFs were subjected to no stress (−), 100 μM SA and 150 μM VRB and stained with G3BP1 and Hoechst (nuclei). Size bar represents 10 μm. E. Depletion of 4E-BP1 inhibits VRB-induced SG formation. WT (no siRNA), control siRNA- (ctrl), 4E-BP1- and 4E-BP1/PERK-treated U2OS cells subjected to 100 μM SA and 150 μM VRB, stained with G3BP1 and % of SG-positive cells was determined. Standard statistics was applied (unpaired Student's t-test (p-values are shown, N = 3)).

Fig 4: Schematic representation of nandrolone-antagonizing effects on OXER1-initiated pathwaw in BC cells.The binding of nandrolone to OXER1 antagonizes the PI3K/Akt/NF-κB pathway leading to a significant decrease of c-Rel nuclear translocation through a strong decline in RACK1expression, which inhibits cell proliferation and mi-initiated PI3K/Akt/ NF-κB pathway and RACK1 expression in BC cells (see text for more details).

Fig 5: The diagram of a potential signaling pathway that RACK1 regulates the cell proliferation and cell invasion in cervical cancer cells.