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. 2014 Jan 8;9(1):e83391. doi: 10.1371/journal.pone.0083391

Figure 1. Quiescent and activated HSCs express Toll like receptors.

Figure 1

(A) mRNA levels of TLR1-13 were quantified by qRT-PCR in three separate preparations of primary rat qHSCs (day 0) and day 10 transdifferentiated myofibroblasts. Data are expressed as relative level of transcriptional difference (RLTD) to TLR1 mRNA expression (n = 3). (B) Thirty micrograms of whole cell protein extract from three separate preparations of quiescent rat HSCs (culture day 1) or activated myofibroblasts (culture day 10) were separated by SDS-PAGE and immunoblotted for TLR3 and GAPDH. (C) Toll-like receptor 3 was visualised in the cytoplasm of rat qHSCs (ex vivo) culture day 1 (bar represents 75 µm). (D) Quiescent rat HSCs were treated with Poly(I∶C) (1 µg/ml) or IL-1α (2 ng/ml) for up to 24 hours; IL-6 mRNA was measured and normalised to β actin (n = 3). (E) Thirty micrograms of whole cell protein extract from two separate preparations of quiescent HSCs or activated myofibroblasts (culture day 10) were separated by SDS-PAGE and immunoblotted for IRAK1, TRAF6 and GAPDH. (F) Activated rat myofibroblasts were treated with IL-1α (2 ng/ml) for up to 24 hours; IL-6 and TIMP1 mRNA were measured and normalised to β actin (n = 3). (*p<0.05, ***p<0.001).