Figure 2
From: Autophagy Correlates with Maintenance of Salivary Gland Function Following Radiation
Autophagy is not robustly induced at acute time points following targeted head and neck radiation.
The head and neck region of female FVB wild-type mice were irradiated. Parotid tissue was collected at 4, 6 and 8, hours after treatment and representative duplicate samples are shown in each immunoblot. UT: untreated. Data displayed as the mean ± SEM. Treatment groups with the same letters are not significantly different from each other. (A.–E.) Protein lysates were prepared from parotid glands and Western blotting was performed as described in Materials and Methods. Tubulin was used to confirm equal loading. All gel images are cropped to display only the important bands for clarity. (A.) Parotid tissues were treated as stated above and collected at 4–8 hours following radiation for immunoblotting and prepared as described in the materials and methods and membranes were probed for Atg7 (top panel), Atg5 (second panel), short LC3 exposure (third panel), long LC3 exposure (fourth panel) and p62 (fifth panel) with Tubulin (bottom panel) as a loading control. (B.) The graph represents the ratio of LC3-II to the loading control Tubulin, which was quantified using densitometry. n = 2 per time point/treatment. (C.) The graph represents the ratio of Atg7 to the loading control Tubulin, which was quantified using densitometry. p < 0.05, n = 3 per time point/treatment. (D.) The graph represents the ratio of Atg5 to the loading control Tubulin, which was quantified using densitometry. p < 0.05, n = 3 per time point/treatment. (E.) The graph represents the ratio of p62 to the loading control Tubulin, which was quantified using densitometry. n = 2 per time point/treatment. (F.) Representative electron microscopy images of parotid glands of untreated mice (top panels), mice at 8 hours post-radiation (middle panels). AL: autolysosome; AP: autophagosome; M: mitochondria; N: nucleus. (G.) Representative images of immunohistochemistry performed using an antibody against Atg5. (H.) Representative images of immunohistochemistry performed using an antibody against LC3.
