Figure 4.

Recruitment of autophagy-related proteins to C. albicans entry sites is associated with plasma membrane damage.

(a) Representative images of C. albicans-infected HeLa cells at 4 h postinfection. Samples were processed for C. albicans (red) and Annexin V (green) staining. (b) Quantification of the percentage of Annexin V + C. albicans invasion sites at 4 h postinfection. Data are mean ± SEM of six independent experiments. (c) Representative images of C. albicans-infected GFP-LC3 HeLa cells at 4 h postinfection. Samples were processed for ALIX (red) and GFP-LC3 (green) staining. (d) Quantification of the percentage of LC3+, ALIX+, or double positive C. albicans invasion sites at 4 h postinfection. Data are mean ± SEM of three independent experiments. (e) Representative images of C. albicans-infected HeLa cells at 4 h postinfection. Samples were processed for C. albicans (blue) and Galectin 3 (red) staining. (f) Quantification of the percentage of Galectin 3 + C. albicans invasion sites at 4 h postinfection. Data are mean ± SEM of three independent experiments. (g) Ultrastructural analysis by transmission electron microscopy of HeLa cells infected with C. albicans for 4 h, showing the presence of plasma membrane damage (indicated by white arrowheads) along the C. albicans invasion sites. Black squares indicate the two inset areas.