Fig. 4.

GAPDH regulates TNFα post-transcriptional regulation. a GAPDH was immunoprecipitated from untreated and LPS-treated (6 and 24 h, 100 ng/mL) BMDMs, and TNFα mRNA presence in IP relative to IgG control assessed via qPCR. (mean + SEM, n = 3). b TNFα mRNA expression measured by qPCR and TNFα protein measured by ELISA in BMDMs treated with LPS (100 ng/mL) over time. c TNFα protein measured by ELISA in BMDMs treated with LPS (100 ng/mL). Cells were washed post 4 h and post 8 h LPS treatment, and supernatants replaced (without LPS) and harvested 20 and 16 h later, respectively (mean + SEM, n = 3). d GAPDH expression analysed by western blotting in LPS-treated BMDMs. e LPS-treated (100 ng/mL, 24 h) BMDMs were pre-treated with HA (10 µM) and GAPDH immunoprecipitated. Bound TNFα mRNA was assessed by qPCR. Data shown are representative of three independent experiments. Unpaired t-test, *p < 0.05; ***p < 0.01