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. 2015 Jan 28;10(12):2158–2170. doi: 10.4161/auto.36439

Figure 4.

Figure 4.

PHF23 associates with LRSAM1 via the PHD domain. (A) HEK293 cells were cotransfected with GFP-PHF23 and LRSAM1-MYC-FLAG for 24 h. Total cell extracts were subjected to IP using either an anti-GFP or a control IgG. LRSAM1 was detected in the immunoprecipitated proteins by western blot. (B) HEK293 cells were treated as in (A), except that cell lysates were immunoprecipitated with an anti-FLAG antibody. PHF23 was detected in the immunoprecipitated proteins by protein gel blot. (C) HEK293 cells were cotransfected with plasmids expressing PHF23PHDΔ and LRSAM1-MYC-FLAG for 24 h. Total cell extracts were subjected to IP using either an anti-FLAG or a control IgG, as indicated. PHF23PHDΔ was not detected in the immunoprecipitated proteins, by western blot. (D and E) GST-PHF23 or GST-PHF23PHDΔ fusion protein and the GST protein immobilized on Glutathione-Sepharose beads were incubated with LRSAM1-FLAG-transfected HEK293 cell lysates at 4°C for 4 h. LRSAM1 and GST were detected in the washed beads, by protein gel blot.