Fig. 5

Increased CLSTN1 localization to cell-cell contacts in tumor-astrocyte co-cultures. (A) IFA analysis of CLSTN1 expression in UW228-LA-EGFP-mCherry-Nuc cells. Red: CLSTN1, green: actin (LA-EGFP), blue: Nuclei (mCherry-Nuc). Arrows indicate CLSTN1 localization at the cell cortex, and arrowheads in cell-cell contacts. (B) IFA analysis of CLSTN1 expression in UW228-LA-EGFP-mCherry-Nuc co-cultured with iNHAs. iNHAs do not express LA-EGFP or mCherry-Nuc. Red dotted line indicates the circumference of iNHA cell clusters, and arrowheads indicate CLSTN1-positive cell-cell contacts. Magnifications of zones indicated with white rectangles show homotypic (a) and heterotypic (b) cell-cell contatcs. (C) Plot of the % total grey values of a 10 µm long line across all detected cell-cell contacts in representative images. Mean and 95% confidence intervals of measured values at indicated positions are shown. Monoculture: N = 17, co-culture: N = 31. P values were calculated using multiple unpaired t-tests. (D) IB analysis and quantification of CLSTN1 expression in primary murine cerebellar astrocytes and UW228 MB cells.