Fig. 4: NSD2 is recruited to and contributes to SRC-3 stabilization.

a Silver staining of LP-1 cells infected with lentivirus carrying SRC-3-3 × flag for 72 h (n = 3 biologically independent experiments). b Co-immunoprecipitation (Co-IP) assay shows interactions between CBP, p300, PRMT1, PRMT4, NSD2 and SRC-3. Input, 2% lysate (n = 3 biologically independent experiments). IP, M2-flag antibody. c Western blotting shows the alteration of SRC-3, NSD2, p300, PRMT4, H3K36me2 protein levels in the wild type (WT) and bortezomib (BTZ)-resistant (BR) MM.1S and LP-1 cells (n = 3 biologically independent experiments). d Proximity ligation assay (PLA) shows protein–protein interaction between NSD2 and SRC-3 (n = 30 cells from 3 biologically independent experiments). e Degradation rate of SRC-3 protein in WT and bortezomib (BTZ)-resistant (BR)-myeloma cells treated with 20 μM cycloheximide (CHX) for up to 10 h (n = 3 biologically independent experiments). f Degradation rate of SRC-3 protein in MM.1S and LP-1 cells infected with lentivirus carrying NSD2-HA (NSD2-OE) or vector control (Vector) for 72 h treated with 20 μM CHX for up to 10 h (n = 3 biologically independent experiments). g SRC-3-3 × flag levels in HEK293T cells co-transfected with full-length NSD2 (FL), SET domain depletion NSD2 (ΔSET), or Y1179A mutation NSD2 (Y1179A) for 48 h (n = 3 biologically independent experiments). h Immunoprecipitation assay for full-length SRC-3 (FL) or AD2-domain depletion SRC-3 (ΔAD2) in HEK293T cells (n = 3 biologically independent experiments). i Immunoprecipitation assay for full-length NSD2 (FL) or truncations with PWWP domain depletion (ΔPWWP), HGM domain depletion (ΔHGM), or PHD domain depletion (ΔPHD) with SRC-3 in HEK293T cells (n = 3 biologically independent experiments). Input, 2% whole lysate; IP, M2-flag antibody or HA antibody. Source data are provided as a Source Data file.