ZNF198 Stabilizes the LSD1–CoREST–HDAC1 Complex on Chromatin through Its MYM-Type Zinc Fingers (original) (raw)
Figure 5
ZNF198-like proteins regulate the chromatin association of LSD1.
(A) HeLa Tet-on cells were transfected with the indicated siRNAs: Luc (firefly luciferase), MYM (ZNF198, ZNF261, and ZNF262), or LSD1. Lysates from these RNAi cells were immunoprecipitated with either anti-ZNF198 (top panel) or anti-LSD1 (middle panel). The IPs and the lysates (bottom panel) were blotted with the indicated antibodies. (B) After RNAi of the indicated proteins, nuclear pellets were generated by subjecting HeLa Tet-on cells to hypotonic lysis followed by centrifugation. Normalized samples from each step were subjected to SDS-PAGE and blotted with the indicated antibodies. (C) Nuclear pellets in lanes 7–9 in (B) were subjected to extraction with a high-salt buffer. Normalized samples from each step were blotted with the indicated antibodies. (D) HeLa cells transfected with siLuc were subjected to fractionation as in B. The nuclear pellet in lane 7 in (B) was digested with micrococcal nuclease (MNase) followed by extraction with 2 mM EDTA. Supernatants (S) and pellets (P) were blotted with the indicated antibodies.