Extended Data Fig. 3: Nalcn deletion does not affect cell proliferation, apoptosis, immune-infiltration, vasculature or ASMA expression in primary tumours in P1^KP, V1^KP or Pdx1^KP mice.

(a) HALO-quantification of Nalcn mRNA transcripts per cell detected by RNA-scope analysis in mouse gastric (GAC), intestinal (IAC) and pancreatic (PAC) adenocarcinomas of the indicated Nalcn genotype (bar=median; *p = 0.0294; ***p = 0.0004; ****=p < 0.0001, two-tailed Mann-Whitney test). Data are tumour fields (5–8 images per tumour) from n = 5 tumours for each Nalcn genotype of P1^KP-GAC, V1^KP-IAC and Pdx1^KP-PAC mice (total n = 45 unique tumours, 289 unique tumour fields). (b) Representative photomicrographs of Nalcn RNA in situ hybridization in GACs (n = 15 biologically distinct tumours, 100 tumour fields) of the indicated Nalcn genotype (scale=50 μm). (c) Left in each is HALO-quantification (Data are mean ± SD) of immunohistochemically-detected tumour cell expression of MKI67 (proliferation), cleaved Caspase-3 (CC3; apoptosis), CD45 (immune cell infiltration), CD31 (endothelial cells) and alpha-smooth muscle actin ASMA; stroma) in five complete biologically independent tumour fields for each Nalcn genotype of P1^KP-GAC, V1^KP-IAC and Pdx1^KP-PAC mice (total n = 45 unique tumours). Two-tailed Mann-Whitney U tests revealed no significant difference in these markers among tumours with different Nalcn genotypes. P-values GAC, IAC, PAC of Nalcn^+/+ vs Nalcn^+/Flx, Nalcn^+/+ vs Nalcn^Flx/Flx, respectively: KI67 (0.4206, 0.4206, 0.4206, 0.5476, 0.2222, 0.5476), CC3 (0.9999, 0.5476, 0.0952, 0.5476, 0.9999, 0.2222), CD45(0.6905, 0.8413, 0.1508, 0.3095, 0.6905, 0.8413), ASMA(0.0556, 0.8413, 0.3095, 0.0556, 0.2222, 0.1508), CD31(0.9999, 0.0952, 0.0952, 0.4206, 0.8413, 0.1508). Right in each are exemplar photomicrographs of the indicated marker in the indicated tumour type (scale=50um).

Source data