FIGURE 6.

Knockdown of PRMT5 methyltransferase and Runx2/p57 expression during mesenchymal cell lineage commitment. C2C12 cells were differentiated to the osteoblast (B) or myoblast (C and D) lineage as described in Fig. 1. pC cells were infected with lentiviral particles coding for shRNAs against PRMT5. Effective down-regulation was confirmed by Western blot analyses 72 h post infection. TFIIB protein levels were used as loading control (A). Runx2-II/p57 and Osteocalcin (Oc/Bglap) mRNA levels were measured as described in Fig. 4, B and C. ChIP assays were performed using antibodies against PRMT5 protein or against the H4R3Me2-symmetrical histone mark (D). Results and statistics are expressed as described in Fig. 4. ***, p < 0.001; ns = non-significant differences.