Extended Data Fig. 1: Fresh cell transplantation and lineage tracing for gene activation from basal and luminal cells.
From: ERG-driven prostate cancer initiation is cell-context dependent and requires KMT2A and DOT1L
(a) Quality control for freshly isolated basal and luminal-derived cells generated in Fig. 1a. (Top) Post-sort analysis of sorted basal and luminal populations. (Bottom) Assessment of Cre recombination efficiency at 5 days post Cre. GFP was used as a surrogate for ERG expression. (b) Images showing prostates harvested at 5 months endpoint. Visible grafts are highlighted in yellow circles. (c) Weights of indicated orthografts harvested at the 5 months endpoint (n = 6 mice per group). (d) Flow cytometry assessing the cell type specificity of K5- and K8-CreERT2. Whole prostates were harvested for analysis. (e) Schematic summarizing that lineage tracing using K5-CreERT2 and K14-CreER to activate EP leads to a severe skin disease and premature death. (f-g) Survival curve (f) and skin histology (g) from indicated mice. Scale bar, 100 µm. (h-j) Mice at harvest (h), survival curve (n = 6 mice per group) (i) and skin histology (j) of indicated mice. Scale bar, 100 µm. (k) IF imaging assessing the cell type specificity and infection efficiency of the indicated Ad-Cre using the Rosa26-YFPLSL/LSL allele (Y) at the 1-week time point (n = 41 sections from 4 mice for K5-Cre group; n = 27 sections from 4 mice for K8-Cre group). Scale bars, 10 µm. (l) Sagittal view of whole prostates with ERG IHC. Two injection sites are shown. Scale bar, 2 mm. Data represent mean ± s.d.; ns, not significant. unpaired two-tailed t-test (c); Log-rank test (f, i).
