Figure 4

Effects of siRNA-mediated R-spondin 3 (RSPO3) silencing on EPS-induced expressions of other myokines in human myotubes. EPS (1 Hz, 4-ms, 20 V/25 mm) was applied to the differentiated human myotubes treated with siRNAs as described in the Methods. Total RNA samples were prepared and subjected to RT-PCR analysis to evaluate mRNA levels for RSPO3 (A), IL-6 (B), CXCL1 (C) and IL-8 (D). Data normalized using RPLP0 transcript were averaged over 3 independent experiments. Multi factor ANOVA with Tukey’s multiple comparison test was used for statistical analyses; the significance of the effect of EPS is denoted as *P < 0.05; the effect of sRNAs compared to the scramble siRNAs control is denoted as # < 0.05. Note that RSPO3 expression is effectively reduced by RSPO3 siRNAs but not by scramble control siRNAs.