Fig 2: The protein expression of MyD88, p-ERK1/2, and p-p38 was detected using immunohistochemistry (×200). Control (A), model (B), DXM (C), YLX (D), S. oblata-H (E), S. oblata-M (F), and S. oblata-L (G). Data are expressed as mean ± SEM. **p < 0.01 versus control group; ##p < 0.01; #p < 0.05 versus model group.

Fig 3: Theaflavin-3,3'-digallate (TF3) suppresses the activation of AKT and ERK1/2 during carotid artery ligation–induced neointimal hyperplasia. TF3 (10 mg/kg) or vehicle was intraperitoneally injected after carotid artery ligation. (A) Immunofluorescence staining of p-AKT (red), a-SMA (green), and DAPI (blue) (above) and their enlarged view (below) are shown. (B) Immunofluorescence staining of p-ERK (red), a-SMA (green), and DAPI (blue) (above) and their enlarged view (below) are shown. Scale bars, 100 and 25 µm.

Fig 4: RET activates the RAS/RAF/MEK/ERK/p90RSK/ERa phosphorylation cascades to promote the expression of estrogen target genes. (A) MCF7 cells were infected with lentiviral control shRNA (shCTL) or shRNA specifically targeting RET (shRET) followed by RNA-seq analysis. Genes positively and negatively regulated by RET are shown by a pie chart. (B, C) Heat map (B) and box blot (C) representation of the expression (FPKM, log2) for genes regulated by RET (unpaired Student's t-test, two-tailed). (D) Hallmark gene set enrichment analysis for genes positively regulated by RET. (E) The overlap of genes induced by E2 and genes positively regulated by RET is shown by a Venn diagram. (F, G) UCSC Genome browser views of the expression of CCND1 and MYC from RNA-seq are shown. Blue, shCTL; red, shRET. (H) MCF7 cells infected with control shRNA (shCTL) or two independent shRNAs specifically targeting RET (shRET#1 and shRET#2) were maintained in the presence of estrogen (E2, 10-7 M, 6 h) followed by RT–qPCR analysis to examine selected estrogen-induced genes. Statistical significance is shown in Supplementary Table S2 (± SEM, *P <0.05; **P <0.01; ***P <0.001). (I) MCF7 cells pre-treated with LOXO-292 (1 µM), BLU-667 (1 µM) or AST487 (1 µM) were then treated with estrogen (E2, 10-7 M, 6 h) followed by RT–qPCR analysis to examine selected estrogen-induced genes. Statistical significance is shown in Supplementary Table S2 (± SEM, *P <0.05; **P <0.01; ***P <0.001). (J, K) MCF7 cells as described in (H) or (I) were subjected to immunoblotting (IB) analysis using antibodies as indicated. Molecular weight is indicated on the right (in kDa).

Fig 5: RAF1 involved in estradiol secretions by activating ERK phosphorylation. The primary ovarian GCs were treated with RAF709, 5 nM for 6 h then induced by FSH (100 ng/mL) for 24 h. (A-D) The protein expression ratios of FSHR, RAF1, ERK-P, and CYP19A1 were respectively detected by WB, and the protein ratios were analyzed in treatment groups relative to the GAPDH protein abundance. (E) Estradiol content was measured in each treatment group by RIA. The values are the means ± SEM of three independent experiments. Different letters indicate a significant difference between the compared groups (P<0.05). Different letters (a, b, c, and d) between two bars show a signi?cant difference (a versus b, b versus c, and c versus d: P <0.05).