Fig 2: Mean Pixel Intensity (MPI) for green (Integrin-ß1) and red (Connexin-32) channels.The MPI was calculated from n = 3 independent experiments per substrate investigated, imaging and analysing n = 3 different DAPI-positive areas in each sample (randomly selected). Different letters indicate significant differences between groups (one-way ANOVA, p < 0.05): lowercase letters are referred to ANOVA results of green MPI, while capital letters denote ANOVA results of red MPI. Note that the green and red MPI levels cannot be compared due to differences in the laser power used and in the fluorophore excitation/emission efficiencies.

Fig 3: IR transiently downregulates expression of genes involved in saliva secretion. (A-D) Immunofluorescent staining shows colocalization of M3R+ (red) with MIST1+ (green) acinar cells (A) on control. M3R expression does not change at 3 h (B), 24 h (C) or 48 h (D) after IR using the Cs source. (E-H) AQP5 (red) localization at the apical surface of MIST1+ (green) acinar cells in control (E) does not change at 3 h (F), 24 h (G) or 48 h (H) post-IR. Scale bars: 100µm. (I) Expression of Mist1 mRNA was significantly reduced at 3 h post-IR (**P=0.002), but recovered by 24 h post-IR (P>0.1). (J) Relative expression of M3r mRNA did not change post-IR using the Cs irradiator (P>0.1). (K) Aqp5 mRNA expression was decreased at 3 h post-IR (*P=0.023), but recovered by 24 h post-IR (P>0.05). (L) Western blot of MIST1, AQP5, CX32, E-CAD, GAPDH and ß-actin proteins in control and irradiated SMGs using the Cs source (n=3). (M,N) Densitometric analysis of MIST1 (M) and CX32 (N) MIST1: *P=0.041; CX32: **P=0.003. (O) Densitometric analysis of AQP5 protein (*P<0.05). (P) Densitometric analysis of E-CAD protein (one-way ANOVA; F=0.215, P=0.883). (Q) Following IR using SARRP, relative expression of Mist1 mRNA was significantly decreased in both irradiated and CL SMGs at 3 h (***P<0.001, *P<0.05, respectively), at 24 h (***P<0.001, *P<0.05, respectively) and at 48 h (*P<0.05) versus control. Mist1 expression was significantly lower in the IR-treated SMGs compared to the CL SMGs (##P<0.01). (R) IR using SARRP also decreased the relative expression of M3r mRNA in both irradiated and CL SMGs at 3 h (**P<0.001, *P<0.01, respectively) versus non-irradiated control. (S) Following IR using SARRP, relative expression of Aqp5 mRNA was decreased in both irradiated and CL SMGs versus control at 3 h (***P<0.001, **P<0.01, respectively), 24 h (**P<0.01, *P<0.05, respectively) and 48 h (**P<0.01, **P<0.01, respectively). Aqp5 expression was significantly lower in the IR-treated SMG compared to the CL SMG (#P<0.05). Error bars indicate mean±s.e.m. Statistical analysis was performed compared to control (non-irradiated) using one-way ANOVA with Dunnett's post-hoc test, or comparing irradiated and CL SMGs using two-way ANOVA with Bonferroni test (n=3-5 each group).