Fig. 3

CEBPD knockdown blunted IL-6 receptor protein expression and signaling in ASM. A Representative immunoblot lanes for tubulin and CEBPD expression. Immunoblot band density quantification results for CEBPD (normalized to tubulin under NT siRNA control or CEBPD siRNA control within groups) show that its levels were induced by BUD exposure among NT siRNA-transfected samples, an effect that was substantially reduced with CEBPD siRNA transfection. B Representative immunoblot lanes for tubulin and IL-6R expression. Immunoblot band density quantification results for IL-6R (normalized to tubulin under NT siRNA control or CEBPD siRNA control within groups) were significantly increased with BUD versus control exposure among NT siRNA-transfected cells, an effect that not was significant for BUD versus control exposures with CEBPD knockdown. C Representative immunoblot results examining pSTAT3 normalized to total STAT3 expression under NT siRNA control or CEBPD siRNA control within groups. pSTAT3 was induced with IL-6 exposure and further augmented with addition of BUD exposure in NT siRNA transfected cells. The augmented pSTAT3 induced by BUD + IL-6 versus IL-6 was no longer significant with CEBPD knockdown. Barplots of signal ratios are of height equivalent to the mean across donors, and the error bars represent standard errors (SEs) across replicates with N = 6 donors per condition. Intact immunoblot bands are provided in Additional file 1: Fig. E11. BUD budesonide, NT non-targeting, pSTAT3 phosphorylated STAT3