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. 2021 Feb 18;12:1140. doi: 10.1038/s41467-021-21355-5

Fig. 5. The collagen-like exosporium protein BclA3 is required for spore entry into intestinal epithelial cells via Fn-α5β1 and Vn-αvβ1.

Fig. 5

a, b C. difficile spores R20291 were incubated with Fn or Vn for 1 h. Samples were processed and visualized for TEM. White arrows indicate anti-Fn or -Vn rabbit antibody and anti-rabbit-gold 12 nm antibody complex. c–e TEM of wild type (ΔpyrE/pyrE+), ΔbclA3, and ΔbclA3/bclA3+ C. difficile R20291 spores. f, h Internalization and g, i adherence in differentiated Caco-2 cells of wild type, ΔbclA3, and ΔbclA3/bclA3+ C. difficile spores preincubated with f, g Fn or h, i Vn. j, l Internalization and k, m adherence in CHO cells ectopically expressing: j α5; k, m β1; or l αv infected with wild type, ΔbclA3, and ΔbclA3/bclA3+ C. difficile spores preincubated with j, k Fn or l, m Vn. Data shows internalization and adherence normalized to wild-type spores. In bars, each dot represents one independent well from three independent experiments. Error bars indicate the mean ± S.E.M. Statistical analysis was performed by two-tailed unpaired Student’s t test, ns, p > 0.05; ***p < 0.001; ****p < 0.0001. a−e Scale bar, 100 nm.