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. 2018 Mar 30;32(9):4941–4954. doi: 10.1096/fj.201701455RR

Figure 6.

Figure 6

Role of p38α in DAMPs-modulated expression of hypertrophy-inducing genes in cardiac fibroblasts. A) Real-time RT-PCR showing time of effect of DAMPs on mRNA expression of Il6, Tgfb1, and Igf1. *P < 0.05, **P < 0.01 (n = 3). B) ELISA showing time of IL-6 secretion from cardiac fibroblasts stimulated with cardiac DAMPs. Red filled circles represent DAMP-stimulated IL-6 secretion, and black-filled squares represent basal secretion without addition of DAMPs (measured up to 6 h only). *P < 0.05 compared with time 0 (n = 3). C) Real-time RT-PCR data showing effect of 10 μM SB203580 or DMSO vehicle control on DAMP-induced expression of hypertrophy-inducing genes after 6 h (n = 9). *P < 0.05, **P < 0.01, ***P < 0.001 (ANOVA with Šidák post hoc test). NS, not significant. Data normalized to Gapdh mRNA levels and expressed relative to the control. D) ELISA showing effect of 10 μM SB203580 or DMSO vehicle control on DAMP-induced IL-6 secretion after 6 h (n = 9). ANOVA with Sĭdák post hoc test. *P < 0.05; NS, not significant. E) Western blot showing DAMP-induced phosphorylation (p-) of HSP27 and p38α after 20 min and inhibition by 10 μM SB203580. Total p38α expression was included as the loading control. Blots are representative of 3 separate experiments. F) Control or Fb-p38α KO mice were injected with tamoxifen, and miniosmotic pumps were implanted for delivery of saline or ISO as in Fig. 2A. Pumps were removed, and heart tissue was collected 1 wk later. Bar chart shows real-time RT-PCR analysis of cardiac Il6 mRNA levels (means ± sem). Group sizes: control saline (n = 11), control ISO (n = 9), and KO ISO (n = 11). ANOVA with Sĭdák post hoc test: not significant. G) Schematic depicting role of fibroblast p38α in modulating cardiomyocyte hypertrophy.