Figure 6 from Oncogenic Cell Tagging and Single-Cell Transcriptomics Reveal Cell Type–Specific and Time-Resolved Responses to <i>Vhl</i> Inactivation in the Kidney

<p><i>Vhl</i>-null cells exhibit time-dependent proliferation and association with ccRCC-like gene expression. <b>A,</b> Representative dual IHC for tdTomato (brown) and Ki67 (purple) counterstained with hematoxylin in kidneys of KO mice harvested early after recombination. Scale bar, 25 μm. Magnification, ×40. Black arrow, dual-positive cell; red arrow, tdTomato-negative Ki67-positive cell. <b>B,</b> Proportion of tdTomato-positive (top) or tdTomato-negative (bottom) cells that are positive for Ki67 by dual IHC in kidneys of Control and KO mice harvested early and late after recombination (<i>n</i> = 2F, 6M for Control early; <i>n</i> = 4F, 2M for KO early; <i>n</i> = 4F, 5M for Control late; <i>n</i> = 1F, 6M for KO late). Pairwise comparisons by Kruskal–Wallis test with Dunn correction. <b>C,</b> UMAP plot depicting RTE cells from Control and KO mice at the early and late time points. Orange, cells expressing <i>Mki67</i>. <b>D,</b> Proportion of tdTomato-positive (top) or tdTomato-negative (bottom) RTE cells that express <i>Mki67</i> in different conditions. Pairwise comparisons tested by one-way ANOVA with Holm–Šídák correction. <b>E,</b> Proportion of tdTomato-positive cells of different PT identities that express <i>Mki67</i> in different conditions. Pairwise comparisons tested by two-way ANOVA with Holm–Šídák correction. <b>F,</b> Violin plot overlaid with boxplot depicting expression score for genes upregulated in ccRCC cells known to be HIF targets (left) and not known to be HIF targets (right) in tdTomato-positive cells from Control and KO mice harvested at early or late time points. <b>G,</b> Scatter plot depicting changes in mean expression scores for HIF-target (top) and non-HIF-target (bottom) genes specifically upregulated in ccRCC, in tdTomato-positive cells of different PT identities from different conditions when compared with those from Control mice at the early time point. <b>B, D,</b> and <b>E</b>, Median and interquartile range plotted. Only significant (<i>P</i> < 0.05) comparisons shown. <b>C–G,</b> scRNA-seq data shown for <i>n</i> = 3F, 1M mice for tdTomato-positive and tdTomato-negative Control early and Control late samples; <i>n</i> = 2F, 1M mice for tdTomato-negative KO early samples; <i>n</i> = 2F, 2M mice for tdTomato-positive KO early samples; <i>n</i> = 2F, 2M mice for tdTomato-positive and tdTomato-negative KO late samples.</p>