Fig 2.

TcdB primarily colocalizes with CSPG4 on 18Co cells. (A) Confocal image of 18Co cells immunostained for Nectin-3 and treated with JF669-TcdB1 for 30 minutes. The dashed box represents the location of zoomed panels. The main image is presented as a max intensity projection, while the zoomed panels are shown as a single Z-plane. Scale bars, 30 µm and 10 µm (zoom). Arrows indicate locations of colocalized signals. (B) Confocal image of 18Co cells treated with JF669-TcdB1 for 30 minutes and immunostained for CSPG4. The dashed box represents the location of zoomed panels. The main image is presented as a max intensity projection, while the zoomed panels are shown as single Z planes. Scale bars, 30 µm and 10 µm (zoom). Arrows indicate locations of colocalized signals. (C) Manders’ coefficient analyses of receptor signal overlap with TcdB (M1) or TcdB signal overlap with receptors (M2). Points represent individual Manders’ coefficients. The error bar in red represents the mean and standard deviation. ****P < 0.0001, Kruskal-Wallis ANOVA test and corrected for multiple comparisons using Dunn’s post hoc test. N = 5 (D) Pearson correlation coefficient analysis of TcdB colocalization with Nectin-3 or CSPG4. Points represent individual Pearson coefficients. The error bar in red represents the mean and standard deviation. ****P < 0.0001, Mann-Whitney U test. N = 5.