Supplementary Figure 2: Claudin-11 contributes to collective cell migration and invasion.
From: Snail-induced claudin-11 prompts collective migration for tumour progression
(a) A heatmap show the expression of genes related to cell adhesion in SAS-pLKO vs. SAS-shSNAI1 cells. The data was from one experiment. (b) RT-qPCR indicated the expression of cell adhesion molecules and (c) claudins family genes in SAS-pLKO/SAS-shSNAI1 (n = 2 independent experiments). (d) Western blots of claudin-3, -4 and -7 in SAS-pLKO/SAS-shSNAI1 cells. The data was from one experiment. (e) Upper: snapshots from phase-contrast videos of TE1-pLKO/TE1-shCLDN11 cells moving on collagen gels. Scale bars, 100 μm. Lower: directionality of migration (n = 10 cells). (f) Left: representative images of 3D invasion assays. Yellow arrow indicates collectively invasive cells. The y-axis indicates invasion distance and y-axis heights, 60 μm. Right: the relative invasion index (two-sided Student’s t test). (g) MTT assay. The effect of CLDN11 knockdown on viability in SAS cells. The p-value is estimated by ANOVA. (h) Left: representative images of Transwell migration assay of SAS-pLKO/SAS-shCLDN11 cells. Scale bars, 100 μm. Right: quantification of Transwell migration assay (two-sided Student’s t test). (i) Left: schema of the mice experiment. Middle: three representative bioluminescence images of the mice at the end of week 3. Right: quantification of IVIS images (n = 10 mice; two-sided Mann-Whitney U test). (j) Representative images of H&E staining in lymph node samples from mice orthotopic model. Cropped regions are indicated by the yellow rectangle in the overview panels. Red arrows indicate the regions with tumor infiltration. Scale bars, 100 μm. (k) Western blots of claudin-11 in FaDu-WT vs. FaDu-LN cells. WT, wild-type; LN, lymph node. The representative data were from two independent experiments with similar results. (a), (b), (c), (d) and (e) were performed in 2.5D. (f), (g) and (h) were analyzed from three independent experiments (n = 3). Images of (e), (f), (h), and (j) are representative of three independent experiments with similar results. Data represent mean values shown in (b), (c), (g) and (i). Data represent mean ± S.D. shown in (f), (g) and (h). N.D., no detection; N.S., no significance. Uncropped images of all blots are shown in Supplementary Figure 8. Source data is provided in Supplementary Table 11.
