Extended Data Fig. 8: PTEN loss promotes POU2F3 in basal-derived SCLC. Related to Fig. 4.
From: Basal cell of origin resolves neuroendocrine–tuft lineage plasticity in cancer
a, T7 endonuclease assay on transformed RPM and RPMA basal organoids after lentiviral infection of LCV2 with sgCtrl or sgPten. Expected products of digestion with editing are 671 and 239 bp. b, Immunoblot of pAKT (Ser473) and total AKT with HSP90 as loading control on transformed RPM and RPMA basal organoids with LCV2-sgCtrl or -sgPten. c, Quantification of tumour volume (mm^3) over time (weeks) in RPM (top) and RPMA (bottom) sgCtrl (passage 1=orange, solid; passage 2=green, dashed), and sgPten (passage 1=blue, solid; passage 2=purple, dashed, passage 3=purple, dotted), basal organoid allografts. Exact number of tumours quantified indicated in figure. d, IHC of RPM and RPMA basal-organoid-derived tumours infected with LCV2-sgControl (sgCtrl) or sgPten for indicated markers (left). H-score quantification for indicated proteins (right) in RPM and RPMA “Ctrl” (parental and sgCtrl-infected tumours, orange) and “sgPten” tumours (purple) with SCLC-dominant histopathology (> 50% of tumour region analyzed is SCLC). Quantification of NSCLC-dominant tumours (only in RPMA) included on far right (both “Control” and “sgPten” tumours). Exact number of tumours quantified from n = 5–6 mice per genotype indicated in figure. Multi-passage tumours included. e, Representative H&E and IHC for indicated markers in RPM and RPMA sgCtrl and sgPten tumours with SCLC histopathology versus regions of NSCLC histology including adenocarcinoma (Adeno), adeno-squamous carcinoma (Adeno-squamous), or Squamous differentiation. H-score quantification for indicated proteins (right) in RPM and RPMA “Ctrl” (parental and sgControl-infected tumours, orange) and “sgPten” tumours (purple) with SCLC-dominant histopathology (> 50% of tumour region analyzed). Quantification of NSCLC-dominant tumours (only found in RPMA) included on the far right (both “Control” and “sgPten” tumours). Exact number of tumours quantified from n = 2–6 mice per genotype indicated in figure. Multi-passage tumours included. f, Stacked bar chart has average proportions of indicated histopathologies in individual RPM and RPMA control or sgPten-tumours. Exact number of tumours analyzed indicated above bars. Error bars, mean ± SEM. Histopathologies determined via analysis of H&E and NKX2-1, P63, KRT5, and SCLC subtype marker staining. LCNEC is large-cell neuroendocrine carcinoma. For gel source data for (a,b), see Supplementary Fig. 1. All scale bars, 50 μm. All statistical tests are one-way ANOVA tests with Fisher’s LSD multiple comparisons; **** p < 0.0001, ns=not significant= p > 0.05, and other exact p-values indicated in figure.
