Tenascin C upregulates interleukin-6 expression in human cardiac myofibroblasts via toll-like receptor 4

Figure 3 Toll-like receptor 4 expression in human cardiac myofibroblasts. A: Data from RT-PCR array showing abundance of TLR mRNA in a pooled sample of human CMF (n = 3 donors). Data expressed relative to 5 housekeeping genes; B: Taqman RT-PCR analysis of TLR2 and TLR4 mRNA levels in human CMF (n = 4 donors). Note log scale. ^bP < 0.001 (paired t-test); C: Western blot of CMF homogenates from 4 donors probed with anti-TLR4 antibody. Fifteen micrograms protein per lane. Molecular size (kDa) on left. TLR4 = 95 kDa; D1: Immunocytochemical localisation of TLR4 in human CMF using a primary antibody to human TLR4 and a Cy3-conjugated secondary antibody; D2: Effect of pre-absorption of primary antibody with TLR4 peptide (8 μg/mL) prior to immunostaining. Nuclei are labelled with DAPI. Loss of immunostaining confirms specificity of the antibody. Scale bar 50 μm; E: Effect of IL-1α (10 ng/mL, 6 and 24 h) on TLR4 mRNA expression in human CMF (n = 4 donors). Data are expressed as mean ± SEM. ^dP < 0.01 vs vehicle (ANOVA post-hoc). TLR: Toll-like receptor.