Figure 1.
TGF-β2 selectively increases Nox4 mRNA expression in primary human TM cells. Serum-starved confluent cultures of primary TM cells were treated (24 hours) with vehicle (0.1% DMSO) or TGF-β2 (5 ng/mL) and (A) relative fold changes in GAPDH-normalized Nox mRNA and p22phox mRNA isoform expression were quantified by qPCR, as indicated. (B) Time-dependent GAPDH-normalized Nox4 mRNA expression in TM cell cultures assayed in the absence (vehicle) or presence of TGF-β2 (5 ng/mL) treated without (solid circles) or with actinomycin D (1 µM for 1 hour) prior to (solid squares) or at 8 hours post TGF-β2 treatment (open diamonds). The continued presence of TGF-β2 (semi-solid diamonds) in the presence of actinomycin D did not alter Nox4 mRNA degradation. Data shown in A and B are the means ± SD (N = 3–5 replicate wells per condition) from separate experiments. ***P < 0.0001; 1-way ANOVA with Tukey's post hoc analysis.