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. 2022 Dec 6;13:7520. doi: 10.1038/s41467-022-35020-y

Fig. 2. BCCs spontaneously transition towards an LY6D+ state.

Fig. 2

a PCA projection of the seven merged human BCC tumor epithelial samples from Fig. 1b. Arrow indicates presumptive lineage transition. b UMAP plot of scRNA-Seq data from human BCC21 tumor epithelial sample with eight distinct clusters. The circled BCC21-Hu-C5 population, is enriched for indicated markers. c Monocle projection of human BCC21 tumor epithelial sample with the same cluster information from Fig. 2b. Arrow indicates transition from an LY6DNeg to BCC21-Hu-C5. d UMAP plot of scATAC-Seq data from three merged human three BCC tumor samples. e UMAP plot of scATAC-Seq data from human BCC21 tumor epithelial sample with four distinct clusters. f Feature plot of the GeneScoreMatrix for LY6D for human BCC21 scATAC-Seq data as shown in Fig. 2e. Monocle projection overlayed the predicted lineage transition. g scATAC-Seq lineage trajectories for GLI1 and LY6D. h Chromatin accessibility for around LY6D locus for each cluster from human BCC21 scATAC-Seq. LY6D is on the negative strand (genes in blue). Peaks enriched for the scATAC-Seq C1/red cluster are indicated in blue. i Motifs enriched in the open regions of each cluster (red) and motifs involved in the transitions between clusters (blue). j Sorting strategies from tumors or mouse skin used for organoid cultures. k Brightfield images of organoids grown from Ly6d+ or Ly6d− epithelial cells isolated from either mouse BCCs or mouse epidermis. l Quantification of the number of organoids from BCCs and normal epidermis. Each point represents number of organoids from either BCCs (n = 5) or mouse skin (n = 3). m Ly6d expression from Ly6d+ or Ly6d− BCCs directly after sorting or from Ly6d+ or Ly6d− tumor epithelial cells cultured as organoids. Each point represents Ly6d expression level normalized to Gapdh from distinct (n = 3) primary tumors from different mice. n. Flow analysis of Ly6d− BCCs cultured as organoids for 14 days. Length of each scale bar is noted in figure. m Error bars represent mean ±SD. l, m, p values were calculated using unpaired, two-tailed t test. Source data are provided as a Source Data file.