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. 2023 Nov 4;14(11):719. doi: 10.1038/s41419-023-06248-3

Fig. 1. Structural and functional characterization of autophagy in HNC cell lines.

Fig. 1

A Representative Electron Micrographs showing subcellular compartments in two HPV− (UM-SCC-4 and UM-SCC-19) and two HPV+ (UD-SCC-2 and UPCI-SCC-152) cell lines. The panels on the right represent a magnification of the panels on the left. Arrows indicate autophagic vacuoles (AVs). B Boxplots show the number of AVs per cellular profile (n > 19). Box are expressed as means, including Tukey analysis. Statistical analysis was performed using ordinary one-way ANOVA corrected comparing False Discovery Rate (two-stage step-up method of Benjamini, Krieger and Yakuteli). C Western blot shows LC3 proteins levels in three HPV− (UM-SCC-4, UM-SCC-10A and UM-SCC-19) and three HPV+ (UD-SCC-2, UPCI-SCC-90, UPCI-SCC-152) HNC cell lines. Actin was used as loading control. LC3-II/LC3-I ratio is reported. D Histogram shows the densitometric protein levels of LC3-II/LC3-I ratio from figure 1C ± SEM. Unpaired t-test was performed. E Confocal micrographs of HPV− (UM-SCC-19) and HPV+ (UD-SCC-2) cell lines transduced with GFP-RFP-LC3 (Red and Green dots) and treated with 10 nM Bafilomycin A1, 1 µM Torin-1 or vehicle (CTR) for 24 h. The panels on the right represent 5x zoom of boxes on the left panels. Nuclei were stained with DAPI (Blue). F Quantification of colocalized dots for each RFP dot. Each dot represents a different field. Statistical analysis was performed using two-way ANOVA test with Dunnett’s correction (n > 9). Bars are expressed as mean ± SD.