Fig 2: IGF-1 inhibits radiation-induced nuclear translocation of Yap in acinar LRCs. (A) Label-retaining assay was performed in Prkcz-/- and wild-type (C57BL/6 J) mice as described in methods section (n = 3 for all groups). 4 days following radiation, animals were injected with 1 or 4 doses of IGF-1 (5 ug/mouse) 24 hours apart. PGs were collected 24 hours following the last dose of IGF-1 (at day 5 or 8 following radiation). (B–E) Representative images of PGs stained for Yap (Magenta) and EdU (Green). White arrows point at Yap-positive LRCs with nuclear Yap. (F) Quantification of the percentage of Yap-positive acinar LRCs. (G) Quantification of the percentage of nuclear-Yap-positive acinar LRCs. Star represents statistical differences vs untreated group (p < 0.05 by one-way ANOVA with follow up Tukey’s multiple comparisons test).

Fig 3: aPKC? is required for functional restoration of irradiated salivary glands with IGF1. Saliva was collected from wild type (C57BL/6 J) and Prkcz-/- mice at days 3 and 30 following radiation treatment as described in the methods section. A minimum of 9 males and 3 females were used per group for saliva collection at day 3. At day 30, at least 5 animals were used for each treatment group. Values for each mouse strain were normalized to their respective untreated control groups. Statistical differences between groups are represented by a star (p < 0.05, One-way ANOVA followed by Tukey’s multiple comparisons test).

Fig 4: aPKC? inhibits proliferation of LRCs in acinar compartment. (A) Experiment setup. Label-retaining assay was performed in Prkcz-/- and wild-type (C57BL/6 J) mice as described in methods section (n = 3 for all groups). PGs were collected 5 days following radiation treatment for analysis. (B) Percentage of Ki67-positive LRCs in acinar compartment of Prkcz-/- and wild-type mice. Representative images of control and irradiated PGs immunostained for EdU (Green) and Ki67 (Magenta) are shown in panels C–F. Proliferative LRCs are shown with a white arrow. Different letters represent statistical differences between groups (p < 0.05 by one-way ANOVA with follow up Tukey’s multiple comparisons test).

Fig 5: Percentage of T560aPKC?-positive and nuclear-Yap-positive LRCs are maintained chronically following IGF-1 treatment. (A) Label-retaining assay was performed in Prkcz-/- and wild-type (C57BL/6 J) mice as described in methods section (3 mice were used per group). 4 days following radiation, animals were given 4 doses of IGF-1 (5 µg/mouse) 24 hours apart. PGs were collected 30 days after treatment. (B,C) Representative images of PGs stained for aPKC?T560 (Magenta) and EdU (Green). (D–G) Representative images of PGs stained for Yap (Magenta) and EdU (Green). (H) Quantification of the percentage of aPKC?T560-positive acinar LRCs. (I) Quantification of the percentage of nuclear-Yap-positive acinar LRCs. Star represents statistical differences vs untreated group (p < 0.05 by one-way ANOVA with follow up Tukey’s multiple comparisons test).