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. Author manuscript; available in PMC: 2020 Jan 23.
Published in final edited form as: Sci Transl Med. 2019 Jan 23;11(476):eaav4754. doi: 10.1126/scitranslmed.aav4754

Fig. 7. Deletion of CG1 inhibits TASCC formation and fibrosis.

Fig. 7.

(A) Coimmunostaining of KIM-1 and pH3 in kidneys from wild-type or CG1−/− mice 6 weeks after AA-induced injury. Upper panels: Stitched images representing about 25% of the kidney cross-sectional area taken at ×200 magnification. Lower panels: Higher-magnification images from the boxed regions in the stitched (upper) images. Scale bars, 500 μm. (B and C) Quantification of KIM-1 and pH3 staining from whole-kidney section images in (A). n = 4. (D) Serum creatinine from wild-type and CG1−/− mice measured immediately before AA-induced injury and weekly thereafter for 6 weeks. (E) Coimmunostaining for TASCC markers, LC3, mTOR, and LAMP2 in wildtype and CG1−/− kidneys after AA-induced injury. Dashed boxes indicate the high-power (HP) region. Arrows, TASCC structures. Scale bar, 10 μm. (F) Quantification of the staining in (E) using NIS Elements software to measure Pearson’s colocalization correlation. n = 3. (G) Upper panel: Imaging of picrosirius red-stained whole-kidney sections from stitched ×100 images using polarized light microscopy. Lower panel: Higher magnification (dashed box) from the stitched images in the upper panel. Dashed line delineates the kidney cortex and medulla regions that were quantified (papillary regions were omitted from quantification). Scale bars, 1 μm (upper) and 100 μm (lower). (H) Quantification of the positive area of images in (G). n = 4. (I) Representative images of Masson’s trichrome staining of kidneys from wild-type and CG1−/− mice after AA-induced injury. (J to L) Quantification of TGF-β, αSMA, and CTGF in wild-type and CG1 kidneys 6 weeks after AA-induced injury by real-time polymerase chain reaction (values are fold change compared to wild-type vehicle-treated mice). n = 3. *P < 0.05, **P < 0.01. Data are means ± SEM. (F and H) ANOVA–Tukey’s post hoc test. (A to D and I to L) Student’s t test. All scale bars, 10 μm.