Extended Data Fig. 8: Influence of HD5 on cell adhesion, cell viability, TEER, DSS-damage and microvilli generation of epithelial cells.
a, b, Fluorescence microscopy analysis of influence of initial adherence on HeLa, CACO-2 and HT29-MTX cells in the presence of 2 μM HD5 for 30 min (a) and quantification of cell attachment on petri dishes assessed by crystal violet staining 1 h post-seeding (b). F-actin is grey and nuclei are blue (DAPI). The bars represent 1 μm. c, Effects of different concentrations of HD5 on cell viability after 48-hour treatment (n = 5). d, The influence of different concentrations of HD5 on the trans-epithelial electrical resistance (TEER) during the polarization process of CACO-2 cells. e, Fluorescence microscopy analysis of the protective effect of HD5 on intestinal epithelial barrier upon 10% DSS damage. ZO-1 is green, F-actin is red and nuclei are blue (DAPI). The bars represent 100 μm. f, SEM analysis (n = 5) of the microvilli on polarized CACO-2 cells cultured in transwell for 5 days in the absence (control) or presence of 2 μM HD5. The bars represent 1 μm. Data are presented as mean±s.d. (b-d, f). Significance between indicated groups was calculated using a one-way ANOVA (Dunnett’s multiple comparison Test), and indicated as the p value. ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. All results are representative of at least three independent experiments.
