Fig. 6. Nucleated CZCs in P1^KP, V1^KP and Pdx1^KP mice are CTCs.

a, UMAP of SCS profiles of CZCs (n = 1,820) and PBMCs (n = 559). b, Gene set enrichment from 2,086 gene sets in UMAP clusters in a. c, Coimmunofluorescence of CZCs and PBMCs in P1^KP (upper) and V1^KP (lower) mice (ZSG; scale bar, 10 μm). Representative photomicrographs of 22 cells identified across n = 20 blood films assessed from n = 5 tumor-bearing animals. d, Autofluorescence of Pdx1^KP-PAC CZC metastases in whole lung of recipient immunocompromised mouse (upper left; scale bar, 0.5 cm). Other images show H&E (representative image of 3,061 metastases evaluated) or coimmunofluorescence of metastases (representative images of 28 metastases evaluated) of P1^KP-GAC or V1^KP-IAC CZC metastases in recipient mice (scale bar, 50 μm). Single-channel images are shown in Supplementary Fig. 1. e, Total metastases per organ in recipient mice injected with 25,000 CZCs. P1^KP Nalcn^+/Flx PAC, n = 5 mice; P1^KP Nalcn^+/+ GAC, n = 2 mice; P1^KP Nalcn^+/Flx GAC, n = 3 mice; V1^KP Nalcn^+/Flx IAC, n = 2 mice; V1^KP Nalcn^+/+ + GdCl₃ IAC, n = 5 mice. Source data are given in Supplementary Table 19. f, Metastasis-free survival of immunodeficient NOD scid gamma recipient mice injected with different numbers (10,000, 1,000, 100 or 10) of P1^KP GAC or Pdx1^KP PAC CZCs (n = 3 mice for each condition). ***P = 0.0002 Mantel–Cox statistic. Source data are available in Supplementary Table 19.