Fig. 7: Identification of SOX9 phosphorylation sites modified by ERK1/2 and their influence on Cyclin D1 expression, SOX9 nuclear location and acetylation. | Experimental & Molecular Medicine

Fig. 7: Identification of SOX9 phosphorylation sites modified by ERK1/2 and their influence on Cyclin D1 expression, SOX9 nuclear location and acetylation.

From: Sublytic C5b-9 induces glomerular mesangial cell proliferation via ERK1/2-dependent SOX9 phosphorylation and acetylation by enhancing Cyclin D1 in rat Thy-1 nephritis

Fig. 7

a–e Rat GMCs were transfected with HA-tagged pcDNA-SOX9 wild-type (WT) or different phospho-mutant pcDNA-SOX9 plasmids (S64A/E or S181A/E) in the presence or absence of sublytic C5b-9. HA-SOX9 protein was purified, and the SOX9 phosphorylation sites were identified by mass spectrometry (MS, a). Cyclin D1 promoter activity and promoter occupation were determined by luciferase reporter (b) and ChIP-PCR assays (c). Cyclin D1 abundance and nuclear SOX9 levels were tested by IB (d). SOX9 acetylation and its association with KAT7, PCAF, and GCN5 were measured by co-IP assays (e). f Rat GMCs overexpressing the ERK1/2 gene were transfected with pcDNA-SOX9 WT or different phospho-mutant pcDNA-SOX9 plasmids for 48 h. The Cyclin D1 protein level was measured by IB. **p < 0.01 versus the pcDNA3.1 or pcDNA-ERK1/2 + pcDNA3.1 group; Δp < 0.05, ΔΔp < 0.01 versus the WT or pcDNA-ERK1/2 + WT group. Data were represented as the means ± SD (n = 3 in each group in all experiments).

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