Figure 3.

Decrease in smooth muscle associated proteins in NOD mice. Total proteins and RNA were extracted from the lacrimal glands of NOD and BALB/c mice and processed for (A) SDS-PAGE / Western blotting and (C) RT-PCR analysis, respectively. (B) Western blot staining was normalized to REVERT^TM total protein stain showing significant decreases in α-smooth muscle actin and calponin but not caldesmon and e-cadherin protein expression in NOD LGs compared to control BALB/c LGs. (D) RT-PCR was normalized to internal GAPDH controls (representative shown) but no difference in mRNA levels was detected. Immunofluorescent double-staining of healthy BALB/c (E–G) and diseased NOD (H-J) LGs for calponin and E-cadherin show decreased staining of calponin in diseased NOD LGs. Star denotes statistically significant difference compared to control; n = 5 for each group; scale bar represents 50 µm (E–J). For protein blots NOD and BALB/c samples were run and cropped from the same gel and for PCR each gene of interest and control gene (GAPDH) were run and cropped from the same gel; for accurate standardization. Full blots are provided in the supplementary data file.