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. 2021 Mar 4;12:603649. doi: 10.3389/fimmu.2021.603649

Figure 6.

Figure 6

Transgene IL-37 expression reduces the pro-inflammatory response of mHSC and KC. (A) Supernatants of freshly isolated murine HSC were collected on day 5, 7, 9, and 12 during myofibroblast differentiation and tested for spontaneous IL-6 release. (B) Cultured HSC were stimulated with LPS (100 ng/ml) on day 8, and TGFβ (100 pg/ml) on day 9. Cells were harvested on day 12 and supernatant tested for IL-6 or CCl2 and IL-10 (C). (D) Freshly isolated mHSC were cultured overnight and stimulated with LPS (100 ng/ml) the next day. Total RNA was collected 6 h after stimulation and analyzed by qPCR. Fold changes of mRNA expression (relative to control) were calculated using the ΔΔCt-method normalized to Rpl13a gene expression. (E) HSC were stimulated as described in (B) and Icam1 and α-Sma was analyzed in cell lysates (day 12) by western blotting. (F) Freshly isolated KC were stimulated with LPS (0.1 μg/ml). Supernatants were tested for IL-6 24 h after stimulation. HSC or KC were isolated from Wt (open boxes/bars) or IL-37tg mice (closed boxes/bars). n = 3, *p < 0.05, **p < 0.01, ****p < 0.0001.