Fig 2: Effects of HuR-targeting siRNA on CXCL8 protein expression in HBE cells. HBE cells were treated for 24 h prior to harvesting cell supernatants for analysis with ELISA and whole cell lysates for analysis with immunoblotting. To determine the level of HuR protein knock-down membranes were probed with a specific HuR antibody, then were subsequently stripped and re-probed with an antibody to GAPDH to ensure equal protein loading (A; representative of 3 separate experiments). Densitometry analysis with % knock-down relative to HRV+CSE alone is also shown (A; n=3). Cell supernatants were analyzed for CXCL8 protein (B; n=6). Data are presented as mean ± SEM. Lipid denotes transfection reagent alone. NT denotes non-targeting control siRNA. Asterisks denote a significant difference between the specified treatments (***p<0.001).

Fig 3: HuR knockdown affects HRV+CSE-induced, but not basal, CSE or HRV-induced CXCL8 protein expression in HBE cells. HBE cells were treated for 24 h prior to harvesting cell supernatants for analysis with ELISA and whole cell lysates for analysis with immunoblotting. To determine the level of HuR protein knock-down membranes were probed with a specific HuR antibody, then were subsequently stripped and re-probed with an antibody to GAPDH to ensure equal protein loading (A; representative of 3 separate experiments). Cell supernatants were analyzed for CXCL8 protein (B; n=3). Data are presented as mean ± SEM. Lipid denotes transfection reagent alone. NT denotes non-targeting control siRNA. Asterisks denote a significant difference between the specified treatments (*p=0.05, **p<0.01 and ***p<0.001). ns = not significant.