Fig. 6.

GRIP1 phosphorylation does not potentiate its GR corepressor function. a THP1 cells (small hairpin scrambled (shSCR) or shGRIP1) or BMMΦ (WT or GRIP1 KO) were treated ± 10 ng ml⁻¹ LPS and ± Dex for 1 h and the expression of indicated genes was analyzed by RT-qPCR as described in Fig. 1a. “Fold repression” was defined as the quotient of the transcript level at lipopolysaccharide (LPS) over LPS+dexamethasone (Dex) condition and is shown as Tukey box-and-whisker plots (n = 4). b THP1 cells expressing WT or S469A/S487A/S493A/S499A (4A) mGRIP1 were treated as in (a) and fold repression of indicated genes is shown. ns, non-significant. c THP1 MΦ or BMMΦ were treated ± Dex ± LPS for 2 h, as indicated, and the levels of GRIP1, GRIP1 phospho-isoforms, GR, CDK9 and HSP90 were evaluated by immunoblotting (quantified in Supplementary Fig. 9d; full-size blots are shown in Supplementary Fig. 10h). THP1 MΦ (d) or BMMΦ (e) treated with ±Dex ±LPS for 1 h were analyzed for GR, GRIP1, CDK9 and phospho-GRIP1 occupancy by ChIP-qPCR as in Fig. 1f (see Supplementary Fig. 9e for more genes). Shown are mean + SD, n ≥ 3, *P < 0.05, **P < 0.01, ***P < 0.001, **** P < 0.0001; one-way ANOVA with Dunnett’s multiple comparison test