Fig. 1: Deletion of Qk in eye lens cells leads to cataracts.
From: Qki activates Srebp2-mediated cholesterol biosynthesis for maintenance of eye lens transparency
a Representative immunofluorescent stains of lens cells in paraffin-embedded sections of ocular tissue from control (Ctrl) and Qk-iCKO mice for GFP (green) and Qki-5 (red) (scale bar: 50 μm) at P19. DAPI (blue): nuclei. Schematic at the far right depicting the morphological impairment of eye lens structure upon Qki loss in Qk-iCKO mice compared to the control mice. b Quantification of GFP+Qki5+ cells among all GFP+ cells in the eye lens tissue represented in a. n = 3 mice/group. The results are presented as means with standard deviation (SD). p = 0.0000001090. ****p < 0.0001 (two-tailed unpaired t-test). c Representative immunofluorescent stains of lens cells in paraffin-embedded sections of ocular tissue from control and Qk-iCKO mice for GFP (green) and AQP0 (red) (scale bar: 50 μm) at P19. DAPI (blue): nuclei. d Representative images of eyes and eye lenses isolated from Ctrl and Qk-iCKO mice at early (P19) and late (P30) timepoints. Scale bar: 0.5 mm. e RIPA-soluble fractions of protein lysates from the isolated lenses of Ctrl and Qk-iCKO mice at early (P19) and late (P30) timepoints visualized using silver staining. f Immunoblots of lenses isolated from Ctrl (n = 3) and Qk-iCKO (n = 3) mice at P19 and P30 for detection of the proteostatic stress markers heat shock protein 90 (Hsp90) and p62. β-actin and Ponceau S: loading control. g Aggregates extracted from Qk-iCKO lenses (red circle) dissolved in 8 M urea and visualized using Coomassie blue staining. The strong Coomassie blue-stained band (red square) was excised and subjected to mass spectrometric (MS) analysis. The most enriched proteins according to MS analysis were ranked as shown in the table. The image of MS machine (Orbitrap Pro) is from Thermo Fisher Scientific website. h, i Representative images of h Congo Red-stained (scale bar: 200 μm) and i transmission electron microscopy-analyzed (scale bar: 250 nm) Ctrl and Qk-iCKO lenses at P30. All the experiments were replicated three times in the lab.
