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. 2017 May 30;21(11):3087–3099. doi: 10.1111/jcmm.13221

Figure 2.

Figure 2

OSM mediates expression of ISG through the type II gp130/OSMR complex. (A) human dermal fibroblasts (HDF) were treated with 20 ng/ml hOSM, 20 ng/ml hIL‐6 and 500 ng/ml hsIL‐6Rα or 10 ng/ml hLIF for 5 hrs. Whole cellular extracts were analysed by SDS‐PAGE and Western blot for MDA5 and RIG‐I. Tubulin staining served as loading control. Blots shown are representative for three experiments. (B–E) HDF were incubated only with transfection reagent (NT) or transfected with control siRNA (siCtrl) or LIFR siRNA (siLIFR). At day two post‐transfection, cells were stimulated with 20 ng/ml hOSM for indicated time periods. Total RNA was prepared and subjected to reverse transcription. (B) IRF1, (C) DDX58, (D) IFIH1 and (E) LIFR mRNA levels were analysed by qRT‐PCR. Relative mRNA levels were normalized to GAPDH, and fold changes were calculated relative to untreated, NT sample (set to 1). Shown are the means (n = 3) with S.E.M. Statistical significance was assessed by unpaired t‐test. **P ≤ 0.01, versus siCtrl.