TBX3 represses TBX2 under the control of the PRC2 complex in skeletal muscle and rhabdomyosarcoma

a, b The expression construct pEF-TBX3 (TBX3) or pEF empty vector (EV) was transiently transfected into RD, RH28, and RH30 cell lines and assayed by qRT-PCR using primers against TBX3 (a) and TBX2 (b). Error bars, standard errors (S.E.) and ***p？≤？0.001 versus EV. c Protein extracts from the transiently transfected isolates in (a) were used for western blot analysis with antibodies against the indicated proteins. d pEF-TBX3 (TBX3) and pEF (EV) were transiently transfected into RD2 cells and examined by western bot analysis against the indicated proteins. GAPDH was used as the loading control. e, f A short-hairpin RNA construct against TBX2 (shTBX2) or a scrambled control (scr) construct was transiently transfected into RD, RH28, and RH30 cell lines and assayed by qRT-PCR using primers against TBX2 (e) and TBX3 (f). Error bars, S.E. and ***p？≤？0.001 versus EV. TBX3 and TBX2 are inversely correlated in sarcoma. (g) Scatter plot representation of mRNA expression of TBX3 and TBX2 in sarcoma patients from The Cancer Genome Atlas (TCGA) (rs？=？？0.5173, p？<？0.0001, N？=？113). (h) mRNA expression data for TBX3 and TBX2 in sarcoma patients were clustered and downloaded using cBioPortal (N？=？263). Red represents higher expression while blue represents decreased expressio