Figure 2.

Serum impairs GR transactivation in a reversible manner. HeLa cells were cultured in either 10 or 50% serum overnight, then treated with 100 nM Dex for 4 h before RNA purification. Transcripts of four well-characterised GR transactivated endogenous genes FK506 binding protein 5 (FKBP5), metallothionein 1X (MT1X), glucocorticoid-induced leucine zipper (GILZ), and period circadian protein homolog 1 (PER1) were measured by qRT-PCR (all normalised to GAPDH) (A, B, C and D). Cells were either continually cultured in either 10 or 50% serum, or cultured in 50% serum then returned to 10% serum as shown in (E) before treatment with 100 nM Dex for 4 h and transcript analysis for MT1X (F), FKBP5 (G), GILZ (H), and PER1 (I). Experiments were performed in triplicate and repeated three times. Graphs show mean±s.e.m. *P<0.01 and **P<0.001. NS, not significant.