Proliferation of HNSCC cell line UD-SCC-2 cells is dependent on NSD3. a, Western analysis with indicated antibodies of whole cell lysates from wild-type or NSD3 depleted UD-SCC-2 cells as indicated. All data were reproduced from three independent experiments. b, Cell proliferation rates of UD-SCC-2 cells expressing CRISPR-Cas9 and two independent NSD3 sgRNAs or a control sgRNA. Data are represented as means ± s.d. from n = 3 biological independent samples. **p < 0.01, n.s., not significant, two-tailed unpaired Student’s t test. Cell lines were numbered as the following: 1. sgControl, 2. sgNSD3–1, 3. sgNSD3–2. P values between the cells are: p (1 vs. 2) = 0.0046, p (1 vs. 3) = 0.0029, p (2 vs. 3) = 0.3249. c, Western analysis of NSD3-depleted UD-SCC-2 cells complemented with structure-guided NSD3 derivatives. All data were reproduced by three independent experiments. d, Validation of NSD3 antibody specificity. Left panel: transient expression of either vector alone or Flag-tagged NSD3 in HEK 293T cells. Right panel: CRISPR-cas9 mediated knockdown of either control or NSD3 in MDA-MB-231 cells. Whole cells lysates were blotted with indicated antibodies. *, non-specific band. All data were reproduced by three independent experiments. For gel-source data, see Supplementary Fig. 1.