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. 2023 Jan 16;13:1079415. doi: 10.3389/fimmu.2022.1079415

Figure 4.

Figure 4

lTSLP induces pro-inflammatory cytokine expression in PBMCs by triggering STAT5 activation. (A-D) PBMCs from RA patients and healthy controls (HC) were cultured for 24h with or without LPS (100 ng/mL) or lTSLP (100 ng/mL). Relative mRNA levels of Il-1β (A), Il-6 (B), Il-8 (C) and Il-10 (D) were analyzed by RT-qPCR. (E) PBMCs from RA patients were treated 24h with 0.25 µg/ml anti-TSLP neutralizing antibody or an IgG antibody control, and the relative mRNA levels of Il-1β, Il-6, Il-8, and Il-10 were analyzed by RT-qPCR. (F) PBMCs from HC and RA were stimulated with 100 ng/mL LPS in the presence or absence of 100 ng/mL lTSLP for 1h. Western blots to examine the expression level of p-STAT5, normalized to β-actin. Relative p-STAT5 expression was quantified in (G). (H) PBMCs from RA patients were treated with 50 μM STAT5 inhibitor 1h before being stimulated with LPS (100 ng/mL) and lTSLP (100 ng/mL) for RT-PCR detection of Il-1β, Il-6, Il-8, and Il-10 mRNA expression. P values were measured by unpaired two-tailed Student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. ns, no significant. Error bar represent standard deviation.