Fig. 5
From: Flightless-I governs cell fate by recruiting the SUMO isopeptidase SENP3 to distinct HOX genes
FLII regulates RbBP5 sumoylation and osteogenic differentiation of DFCs. a HeLa cells treated with control siRNA or FLII siRNA were transfected with Flag-RbBP5 and SUMO2 as indicated, and SUMOylation of RbBP5 was monitored by anti-SUMO2 immunoblotting following anti-Flag-IP. The asterisk represents the sumoylated RbBP5 species. b, c Total RNA was isolated from DFCs, after treating cells with two independent FLII siRNAs or control siRNA. Expression of FLII (b) and DLX3 mRNA (c) was determined by RT-qPCR. Values (normalized for GAPDH expression) represent the mean of three experiments ± SEM. d Cell lysates originating from DFCs (b, c) were separated by SDS-PAGE and analyzed by immunoblotting with the indicated antibodies. e ALP activity was quantified in differentiating DFCs, 10 days after transfection with FLII siRNAs or control siRNA and incubation with osteogenic differentiation medium (ODM). f Mineral deposits in DFC cultures were measured by alizarin red staining after 5 weeks of culture with ODM and transfection with two independent FLII siRNAs or a control siRNA