Fig. 2: HDAC1 deacetylates and stabilizes HP1γ protein.
From: Deacetylation induced nuclear condensation of HP1γ promotes multiple myeloma drug resistance
a Silver staining for flag-pulldown of flag-HP1γ complex in HEK293T cells. b Co-IP assay to show interactions between endogenous HP1γ with HDAC1 and (c) endogenous HDAC1 with HP1γ in MM.1 S cells. Input, 2% whole cell lysate. d Coomassie blue staining to show in vitro expression of GST-fusion HP1γ protein and in vivo expression of flag-HDAC1 protein in HEK293T. e GST-pulldown assays to show the interaction between purified HDAC1-flag and GST-tagged-HP1γ protein in vitro. Input, purified HDAC1-flag. f Levels of HP1γ protein in HEK293T cells with increasing overexpression of HDAC1 by western blotting. g Western blotting shows the alteration of HP1γ, HDAC1 and HDAC2 levels in WT and BR MM.1 S and LP-1 cells. h Silver staining for flag-pulldown in the WT and BR LP-1 cells infected with lentivirus carrying HP1γ-flag for 72 h. i Alteration of interaction between HP1γ and HDAC1 in WT and BR LP-1 and MM.1 S cells infected with HP1γ-flag for 72 h. Input, 2% lysate. j Degradation rate of HP1γ protein in WT and BR MM.1 S and LP-1 cells treated with 20 μM cycloheximide (CHX). k Western blotting shows the HP1γ protein level in the HEK293T cells transfected with flag-tagged HP1γ WT, K5R and K5Q. l Ubiquitination status of HP1γ-WT-flag, HP1γ-K5R-flag or HP1γ-K5Q-flag in HEK293T cells. m Polyubiquitination linkage features of HP1γ-flag mapping by positive mutations of ubiquitin in HEK293T cells. n K11-linked polyubiquitination of HP1γ-flag in HEK293T cells. Source data are provided as a Source data file.
