Clinically relevant temperature change-induced expression of miRNAs alters the PKCα protein levels. (A) Human SAECs were incubated at 32°C, 37°C, or 39.5°C for 24 h and cell lysates were immunoblotted for PKCα. Representative of four similar blots and quantified band densities for PKCα normalized to β-tubulin and expressed as percent of levels in 37°C cells. (B) HEK 293T cells were transfected either with a combination of miRNA inhibitors against hsa-miR-27b-5p, hsa-miR-1260a, and hsa-miR-92a-1-5p or an equal concentration of a non-targeting scrambled-sequence inhibitor control. Transfected and untransfected cells were incubated at 32°C, 37°C, or 39.5°C for 24 h and immunoblotted for PKCα. A representative immunoblot and quantified band densities from four similar blots normalized to ß-tubulin and expressed as percent of levels in untreated 37°C cells are shown. Mean ± SEM. (*) P < 0.05, (†) P < 0.04 versus untreated 37°C and 39.5°C cells and miRNA inhibitor-treated 32°C cells, (¶) P < 0.02 versus untreated 39.5°C cells; n = 4.
