Extended Data Fig. 3: The role of SPP1 in human PDAC development.
From: SPP1 is required for maintaining mesenchymal cell fate in pancreatic cancer
a, RT-qPCR analysis of the expression of epithelial markers KRT19, CDH1, and EPCAM, and mesenchymal markers FN1, S100A4, and VIM between Human PDAC; SPP1wt/wt and Human PDAC; SPP1Δ/Δ organoids (n = 3 biological replicates), normalised using SPP1wt/wt organoids values. b, Validation of CRISPR/Cas9 SPP1 knockout by Western blot. n = 3 independent experiments. Gel source data are provided in Supplementary Fig. 1. c, Image of the subcutaneous tumours formed by Human PDAC; SPP1wt/wt and Human PDAC; SPP1Δ/Δ organoids. Scale bar length is 0.5 cm. d, The sizes of subcutaneous tumours of (c) (n = 6 mice). e, Immunofluorescence staining for KRT19 (green), and VIMENTIN (red) of subcutaneous tumours from (c). Scale bar is 50um. f, Comparison of the percentages of different cancer cell subpopulations of (e) (n = 5 mice). g, h, H&E and immunohistochemistry for KRT19 of subcutaneous tumours from (c). n = 3 mice. Scale bar: 2 mm. i, RT-qPCR analysis of the expression of epithelial markers KRT19, CDH1, and EPCAM, and mesenchymal markers FN1, S100A4, and VIM between Human PDAC; Plv-con and Human PDAC; SPP1 overexpression organoids (n = 3 biological replicates), normalised using Plv-con organoids values. j, Image of the subcutaneous tumours formed by Human PDAC; Plv-con and Human PDAC; SPP1 overexpression organoids. Scale bar length is 0.5 cm. k, The sizes of subcutaneous tumours of (j) (n = 6 mice). l, Immunofluorescence staining for KRT19 (green), and VIMENTIN (red) of subcutaneous tumours from (j). Scale bar is 50um. m, Comparison of the percentages of different cancer cell subpopulations of (l) (n = 5 mice). n, o, H&E and immunohistochemistry for KRT19 of subcutaneous tumours from (j). n = 3 mice. Scale bar: 2 mm. For a, d, f, i, k, m data are mean ± s.e.m. P values were calculated using two-sided t-tests.
