Fig 2: α-Br-TMC inhibits both JAK2 and STAT5 phosphorylation.Ba/F3 cells were pre-treated 30 minutes with the indicated concentrations of TSA and α-Br-TMC (A) or with 10 µM α-Br-TMC and DMSO vehicle (B, C) and further stimulated with 5 ng/mL IL-3 for 30 minutes (A), 15 minutes (B) or 5 minutes (C). Final DMSO concentration was 0.02% in (A) and 0.01% in (B, C). Protein whole cell extracts (panels A, C) or cytosolic (C) and nuclear (N) extracts (panel B) were prepared as described in Materials and Methods and analyzed by Western-blot using antibodies specific for phospho-STAT5 (pSTAT5), phospho-JAK2 (pJAK2), STAT5A, STAT5B, STAT5A and B, JAK2 and α-tubulin (loading control and cytosolic-specific marker). SDS-PAGE in (C) was shorter, explaining why the STAT5 mobility patterns (α-Br-TMC-induced shift and STAT5A and B doublet) are not as apparent as on immunoblots in (A, B).

Fig 3: The effects of BSZY-D on activating GATA 1 and CSN5, inducing PRLR deubiquitination and increasing the expressions of JAK2/STAT5 signaling pathway and kisspeptin. Hence, BSZY-D had an efficient role in HPRL infertility improvement.

Fig 4: STAT4 is expressed in neutrophils, is activated by IL-12, and regulates gene expression profile of neutrophils.BM neutrophils were treated with IL-12 (40 ng/mL) for indicated time points. (A) The expression of phosphorylated STAT4/STAT4 (p-STAT4/STAT4) or (B) p-JAK/JAK was examined by Western blot. Representative image from 3 independent experiments (n = 3–5 mice total). (C) BM neutrophils were either pretreated with the JAK2 inhibitor (JAK2i) gandotinib (1 μM) or left untreated, and 15 minutes later, all samples were stimulated with IL-12 for 30 minutes, and expression of p-STAT4/STAT4 was determined by Western blotting at the indicated time points (n = 5 mice/per group in 3 independent experiments). (D) Heatmaps of real-time RT2 Profiler PCR Array showing differential expression of genes between WT and Stat4–/– IL-12–treated and –untreated samples. Differences between groups were assessed by 2-tailed Student’s t test (*P < 0.05), using the RT2 Profiler PCR Array Analysis (n = 3 samples/group). Data are mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001 using 1-way ANOVA followed by Tukey-Kramer post hoc test.

Fig 5: SYQP inhibited the activation of TNF-α and IFN-γ induced inflammatory death. The total proteins were extracted and analyzed by western blot to determine the protein expression levels in lung tissues of ARDS mice. (A,B) The protein expression levels of the phosphorylation of JAK2 and STAT-1. (C,D) The protein expression levels of NLRP3 and Caspase-1. (E,F) The protein expression levels of Caspase-8 and Caspase-3. The bars represent the mean ±SD (n=3); ####p < 0.0001, ###p < 0.001 and ##p < 0.01 versus the control group; ****p < 0.0001, ***p < 0.001, **p < 0.01 and *p < 0.05 versus the model group.