Extended Data Fig. 1: CD9 identification.
From: CD9 identifies pancreatic cancer stem cells and modulates glutamine metabolism to fuel tumour growth
a, YFP stain of pancreatic ducts of R26-LSL-YFP; Ck19-CreER mice 2 weeks post-tamoxifen. Scale bar, 50 μm, inset 50 µm x 50 µm. b, Flow cytometry analysis and genotyping of unrecombined and recombined cells from KFCkY pancreas 2 weeks post-tamoxifen. Expected band sizes (base pairs) are indicated; see Source Data for uncropped gels. c, CD44 and CD133 stain in consecutive pancreatic sections of KFCkY mice 2 weeks post-tamoxifen. Non-responsive regions magnified. Black arrows show apical expression of CD133 in non-responsive cells. Scale bar, 50 µm. d, CD44 gating strategy using rat IgG2b isotype control on KFCkY pancreas 2 weeks post-tamoxifen. Related to Fig. 1d. e, Validation of the CD44 antibody using rat IgG2b isotype control on KFCkY tumours; quantification of CD44+ cells. f, Quantification of transformed/CD44+ YFP+ foci in pancreatic tissues of KFCkY mice two (n = 5) and four (n = 3 biologically independent animals) weeks post-tamoxifen. Dot plot shows mean, s.d. g, Human PDAC markers among upregulated genes in microarray. Fold changes relative to NT cells. h, Flow cytometry analysis of CD44 and CD9 expression on live, YFP+ cells from KFCkY pancreas 4 weeks post-tamoxifen. i, Quantification of live tumour cells (DAPI−YFP+, n = 3 biologically independent animals) from (h). Bar chart shows mean, s.d., p = 0.0013, two-sided t test. j, CD9, E-cadherin and DAPI staining on pancreatic tissues of KFCkY mice 4 weeks post-tamoxifen. Scale bar, 50 µm, 10 μm (inset). k, CD9 gating strategy using rat IgG2a isotype control on live KPCY tumour cells. Related to Fig. 1i. l, Validation of the CD9 antibody using rat IgG2a isotype control in live KPCY PDAC organoids; quantification of CD9+ cells. All flow cytometry plots and immuno-staining data shown in Extended Data Fig. 1 are representative of at least three biologically independent animals/experiments.
