Figure 3. Mutant NSD3-mediated H3K36me2 synthesis promotes oncogenic programming and LUSC tumorigenesis.

a, Representative μCT scans at 90 and 120 days after adenoviral-Cre mediated tumor induction in PSC and PSCN mice. Scale bars: 10 mm. b, Quantitation of tumor volume as in (a) at 120 days (n = 5 mice for each group). c, Representative HE and IHC staining of lung tissue from PSCN and control PSC mutant mice (n = 8 mice for each group). Scale bars: 100 μm. d, Quantification of proliferation (Ki67⁺-cells) as in (c). P values determined by two-tailed unpaired t-test. (b, d). e, Kaplan-Meier survival curves of PSCN (n = 11, median survival = 145 days) and PSC (same data as in Fig. 1f, n = 10, median survival = 200.5 days) mutant mice, indicated P value determined by log-rank test. f, Western blots with the indicated antibodies of LUSC tumor lysates from PSCN and PSC mice. Two independent and representative samples are shown for each genotype. g, Western blots with the indicated antibodies of cell lysates from either control (sgControl) or NSD3-depleted (sgNSD3) PSC_N cells. H3 and tubulin shown as loading controls in (f, g). h, GSEA identifies upregulation of hallmark gene sets: MYC targets and mTOR signaling (MSigDB: M5928, M5924) in RNA-seq data from PSCN versus PSC tumor biopsies and PSC_N cells ± sgNSD3 (n = 3 biologically independent samples per group). Normalized enrichment scores (NES) and nominal P values are provided (detailed statistics description in Methods). i, CUT&RUN profiles of H3K36me2 (upper panel) or H3K27me3 (lower panel) over averaged gene body for all genes or dDEG in PSC_N cells ± sgNSD3 as indicated. j, Quantification of H3K36me2 and H3K27me3 peak intensity change in PSC_N cells ±sgNSD3 on the indicated gene sets. dDEG n = 234, all genes n = 16091. Two-sided t-test P value provided.