Fig. 7: NALCN loss-of-function increases shedding of ntCZCs.

a, ntCZCs identified in individual nontumor-bearing P1^RNalcn^+/+ (n = 87), P1^RNalcn^+/Flx (n = 50) and P1^RNalcn^Flx/Flx (n = 37) mice. Bar, median. ****P < 0.0001, two-tailed Mann–Whitney U-test. Source data are available in Supplementary Table 13. b, UMAP of 201,183 SCS profiles of PBMCs, tCZCs and ntCZCs as well as cells derived from the indicated normal and malignant mouse tissues. c, Coimmunofluorescence of ntCZCs and PBMCs in peripheral blood smears of P1^RNalcn^Flx/Flx mice (ZSG; scale bar, 10 μm). Representative photomicrographs of 11 cells identified in n = 20 blood films from n = 4 mice. Single-channel images are shown in Supplementary Fig. 1. d–f, Direct ZSG-immunofluorescence photomicrographs of ZSG⁺ cells in lung and kidney (scale bar, 50 μm) (d), and enumerated in lung (no Cre, n = 2 mice, 5 lung lobes; Nalcn^+/+, n = 3 mice, 9 lung lobes; Nalcn^+/Flx, n = 3 mice, 8 lung lobes; Nalcn^Flx/Flx, n = 5 mice, 12 lung lobes; NS P = 0.1312, *P = 0.0168, two-tailed Mann–Whitney U-test) (e) and kidney (no Cre, n = 2 mice, 4 kidney sections; Nalcn^+/+, n = 3 mice, 11 kidney sections; Nalcn^+/Flx, n = 3 mice, 10 kidney sections; Nalcn^Flx/Flx, n = 5 mice, 18 kidney sections; ****P < 0.0001, two-tailed Mann–Whitney U-test) (f). g, Organ heatmap of total numbers of ZSG⁺ cell clusters per mouse identified in organs of recipient mice injected with P1^RNalcn^Flx/Flx ntCZCs. h, Coimmunofluorescence of P1^RNalcn^Flx/Flx ntCZCs (arrows) incorporated into the kidneys of recipient mice (arrows indicated ZSG⁺ cells; scale bar, 50 μm). Representative photomicrograph of n = 5 ZSG rests identified in one tissue field from n = 5 mice. Single-channel images are shown in Supplementary Fig. 1. GLO, glomerulus. i, Confocal laser scanning microscope image of P1^RNalcn^Flx/Flx CZCs incorporated into the renal cortex of recipient mice. Scale bar, 100 μm. Representative image of n = 2 mouse kidneys assessed.

Source data