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. Author manuscript; available in PMC: 2023 Mar 17.
Published in final edited form as: Cell Chem Biol. 2021 Aug 31;29(3):386–397.e9. doi: 10.1016/j.chembiol.2021.08.004

Figure 3. MS9715 degrades NSD3 in cells.

Figure 3.

(A-B) Immunoblotting for NSD3S and GAPDH in EOL-1 (A) and MM1.S (B) cells after a 48-hour treatment with the indicated concentration of MS9715, BI-9321 or MS9715N.

(C-D) Immunoblotting for NSD3S in EOL-1 (C) and MM1.S (D) cells treated with 2.5 μM of MS9715 for the indicated duration.

(E) DC50 and Dmax values of MS9715 in MOLM13 cells, shown as the mean ± SD from three independent experiments. MOLM13 cells were treated with MS9715 for 48 hours. See also Figure S1.

(F-G) Immunoblots for NSD3S using the indicated cells pre-treated with DMSO, 10 nM of MLN9708 (F) or 0.3 μM of MLN4924 (G) for two hours, followed by a 48-hour treatment with 5 μM of MS9715.

(H-I) Immunoblots for NSD3S and GAPDH using EOL-1 (H) and MM1.S (I) cells pre-treated with the indicated concentration of Ac-VHL or Ac-VHL-Me relative to DMSO for two hours, followed by a 48-hour treatment with 5 μM of MS9715.

(J) Immunoblots for NSD3S, VHL and GAPDH after a 48-hour-treatment of 293FT cells, either wildtype (WT; left) or with the CRISPR/Cas9-mediated KO of VHL (right), with the indicated concentration of MS9715, relative to mock.

(K) Immunoblots (IB) for ubiquitin (Ub) after anti-Flag immunoprecipitation (IP) of extracts prepared from 293FT cells expressing Flag-tagged NSD3S, after a 48-hour treatment with the indicated concentration of DMSO, BI-9321 or MS9715.