Figure 2
From: Arginine methyltransferase CARM1/PRMT4 regulates endochondral ossification
CARM1 interacts with and methylates Sox9. (A) Histological analysis of E15.5 wt embryos. Sections of humerus were stained with CARM1 and Col2a1 mRNA probes. Sox9 is stained by ISH and immunohistochemistry (IHC). (B) Sox9 interacts with CARM1 in vitro. Recombinant CARM1 protein and GST-Sox9 fragments were mixed and subjected to a GST-pull down assay. (C) Sox9 recombinant proteins and Histone H3 were incubated with PRMT1 and CARM1 in the presence of [3H] AdoMet. (D) Endogenous Sox9 methylation was detected in mouse primary cultured chondrocytes (left panel). Sox9 multiple point mutants a (mt(a)) and b (mt(b)), wt Sox9, Flag-tagged PABP1 (as a positive control) and Flag-tagged Runx2 (as a negative control) expression plasmids were transfected into 293T cells (right panels). (E) Schematic representation of Sox9 mutants used in the methylation assay.