Extended Data Fig. 10: Knockdown of Claudin 2/10 redirects metastasis to the kidney.
From: Vascular heterogeneity of tight junction Claudins guides organotropic metastasis
(a) Representative H&E images of the kidney of mice in the indicated groups. Scale bar, 50 µm (upper), 25 µm (lower). (b) Percentage of mice with kidney lesion in the indicated groups. LentiCRISPRv2 control vector, n = 15; LentiCRISPRv2 Cldn2 and Cldn10, n = 20 mice. (c) Representative images for GFP immunolabeling in kidney. Scale bar, 50 µm (upper), 25 µm (lower). (d) Number of mice with GFP positive kidney. (e) Quantification of percent of the GFP positive area in the indicated groups, n = 12 mice per group. (f) Orthotopic 4T1 tumor volume over time in the indicated groups. LentiCRISPRv2 control vector without fibrosis, n = 14; LentiCRISPRv2 control vector with fibrosis, n = 11; LentiCRISPRv2 Cldn2 and Cldn10 without fibrosis, n = 12 mice; LentiCRISPRv2 Cldn2 and Cldn10 with fibrosis, n = 10 mice. (g) Number of mice with GFP positive kidney in the indicated groups. LentiCRISPRv2 control vector without fibrosis, n = 14; LentiCRISPRv2 control vector with fibrosis, n = 11; LentiCRISPRv2 Cldn2 and Cldn10 without fibrosis, n = 12 mice; LentiCRISPRv2 Cldn2 and Cldn10 with fibrosis, n = 10 mice. (h) Representative GFP and CD31 staining of the kidneys in mice with Claudin 2 and Claudin 10 down regulation together with or without kidney fibrosis. (i) Representative GFP and CD31 staining in the metastatic kidney with Claudin 2 and Claudin 10 downregulation. For i-j, individual images are shown in Supplementary Fig. 10A-C. Scale bar, 25 µm. (j) Representative images for CD31 and quantification of percent of the CD31 positive area in tumor when downregulate Claudin 2/10 in the kidney. Scale bar, 50 µm. LentiCRISPRv2 control vector, n = 5; LentiCRISPRv2 Cldn2 and Cldn10, n = 7 mice. (k) Representative images for CD31 and quantification of percent of the CD31 positive area in tumor when downregulate Claudin 2 and Claudin 10 with or without kidney fibrosis. Scale bar, 50 µm. LentiCRISPRv2 control vector without fibrosis, n = 5; LentiCRISPRv2 control vector with fibrosis, n = 4; LentiCRISPRv2 Cldn2 and Cldn10 without fibrosis, n = 4 mice; LentiCRISPRv2 Cldn2 and Cldn10 with fibrosis, n = 6 mice. Data are presented as mean values +/− SEM. For b: two-sided Chi-square. For f: two-way ANOVA with Sidak’s multiple comparisons test. For j: Unpaired two-tailed t test, with Welch’s correction applied for unequal variances (determined by F-test). For k: one-way ANOVA with Tukey’s multiple comparisons test. P values are listed, ns: not significant.
