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. 2014 Aug 18;9(8):e105354. doi: 10.1371/journal.pone.0105354

Figure 2. Insulin-dependent decrease of 11beta-HSD2 activity and mRNA is reversible but HSD11B2 half-life is not affected.

Figure 2

(A) 11beta-HSD2 activity was measured in HT-29 cells in presence (black bars) and absence (white bars) of 10−7 M insulin for 24 h. The insulin effect was reversible on washout with PBS 24 h or 48 h of follow-up (hatched bars). (B) HSD11B2 expression was assessed by qRT-PCR in HT-29 pretreated with the protein synthesis inhibitor CHX (10 µM) for 1 h and treated with insulin (10−7 M) for 24 h. Each data point is expressed as a percentage of the control value. (C) HT-29 cells were pretreated with (filled circles) or without (filled rhombus) insulin (10−7 M) for 12 h. The cells were then treated with 25 µM of the mRNA synthesis inhibitor DRB, without or with insulin (10−7 M) (defined as time zero). At the indicated time points thereafter, total RNA was isolated, and the steady state level of HSD11B2 mRNA assessed. Each data point is expressed as a percentage of the maximum determined at time zero.