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. 2019 Jul 31;10:1721. doi: 10.3389/fimmu.2019.01721

Figure 6.

Figure 6

Pellino-1 regulates macrophage STAT-1 signaling. MDMs were transfected with siRNA targeting Pellino-1 or scrambled siRNA (control). (A,B,D) 48 h following transfection, cells were stimulated with IFN-γ [20 ng/ml], IL-4 [20 ng/ml], IL-10 [20 ng/ml] or LPS [1 ng/ml] for 24 h. Whole cell lysates were analyzed by Western blot using antibodies specific to STAT-1, pSTAT-6, Pellino-1 or actin for 4 independent donors (A). pSTAT-1 bands for IFN-γ-treated cells were analyzed by densitometry (B). In separate experiments cells were challenged with NTHi (MOI 1 and 10) for 24 h and lysates probed for Pellino-1 and actin (C). RNA was purified from cell lysates and transcribed to cDNA. CD206 gene product was measured by qPCR (D). Fold changes are shown relative to media treated MDMs transfected with scrambled siRNA (first open bar). Data are presented as mean ± SEM of n = 4 independent experiments. Significant differences are indicated by **P < 0.01.