Fig. 2: LATS1/2 in cancer cells increases the proportion of neural cell adhesion molecule 1 (NCAM1)⁺alpha-smooth muscle actin (αSMA)⁺ CAF subpopulation.

A UMAP plot showing the CAF clusters from WT or LATS1/2 dKO 4T1 breast tumors. B Bar plot showing the percentage of each CAF cluster among total CAFs from WT or LATS1/2 dKO tumors shown in A. C Violin plots showing the differential expression of Ncam1 or Acta2 in each mouse breast CAF cluster. Colors correspond to the indicated CAF cluster in the UMAP plot of CAFs shown in A. D Feature plot showing single cells with Ncam1 and Acta2 co-expression on the UMAP plot of mouse breast CAF clusters. Color threshold corresponds to the expression level of the indicated gene. E WT or LATS1/2 dKO 4T1 breast cancer cells were injected into BALB/c mice, and tumors were subjected to flow cytometry analysis 16 days after transplantation. Representative flow cytometry plots showing the frequency of NCAM1⁺αSMA⁺ CAFs are shown in the left panel. Cells were gated on CD45^–epithelial cell adhesion molecule (EpCAM) ^–CD31^–podoplanin (PDPN)⁺ cells. Data are represented as the mean ± SEM of the percentage of NCAM1⁺αSMA⁺ CAFs in tumors among total CD45^-EpCAM^-CD31^-PDPN⁺ cells; n = 3 samples; pooled 4–11 tumors in each group. *p < 0.05, unpaired two-tailed t-test. F Representative immunofluorescence image of mouse breast tumor tissues. Paraffin-embedded sections of WT 4T1 breast tumors were subjected to immunostaining analysis of NCAM1 (green) and αSMA (magenta) or 4’,6-diamidino-2-phenylindole (DAPI) for DNA (blue). Data are representative of three tumors. Scale bar = 50 μm.