Fig 2: Histological characterization, transcript analysis and uromodulin processing in Kcnj1-/- mice. (a) Haematoxylin and eosin (H&E) staining of 12 weeks Kcnj1+/+ and Kcnj1-/- kidneys. Scale bars: low magnification - 2 mm, high magnification - 100 µm (b) Quantitative PCR analysis of tubular marker genes in Kcnj1-/- kidneys. Glom (Glomerulus): Nphs2 (Podocin); PT (Proximal Tubule): Slc5a2 - Sodium/glucose cotransporter 2 (SGLT2), Aqp1 - Aquaporin 1; TAL (Thick Ascending Limb): Slc12a1 - Na/K/Cl cotransporter 2 (NKCC2), Umod - Uromodulin; DCT (Distal Convoluted Tubule): Slc12a3 - Na/Cl cotransporter (NCC), Pvalb - Parvalbumin; CNT/CD (Connecting Tubule/Collecting Duct): Aqp2 - Aquaporin 2, Scnn1b - Sodium channel epithelial 1 beta subunit (ß-ENaC), Scnn1g - Sodium channel epithelial 1 gamma subunit (?-ENaC); Cldn16 - Claudin 16 and Clcnkb (Cl channel Kb) are localized in both TAL and DCT. (c) Representative Western blot analysis on whole kidney lysates of 12 weeks Kcnj1+/+ and Kcnj1-/- mice. ß-actin was used as a loading control. (d) Representative Western blot analysis on kidney membrane fractions of 12 weeks Kcnj1+/+ and Kcnj1-/- mice. Flotillin-1 was used as a loading control, whereas GAPDH was used to test cytosolic contamination. (e) Representative Western blot analysis on kidney cytosolic fractions of 12 weeks Kcnj1+/+ and Kcnj1-/- mice. GAPDH was used as a loading control, whereas Flotillin-1 was used to test membrane contamination. (f) Immunofluorescence analysis for UMOD (green) and NKCC2 (red) in Kcnj1+/+ and Kcnj1-/- serial kidney sections. Scale bars: low magnification - 50 µm, high magnification - 15 µm. Full length blot images can be found in Supplementary Figure S2.

Fig 3: Western blot showing YAP/TAZ expression among HCC tissue. Lysates of HCC tissue were analyzed by Western blotting with anti-YAP/TAZ and anti-GAPDH (control) antibodies. For the original Western blots, see Supplementary Figure S1.

Fig 4: Effect of chronic exposure to ozone on 78-kDa glucose-regulated protein (GRP78) expression in rat hippocampus. (A) Shows the densitometry analysis of the protein levels for GRP78 from rat hippocampus following exposure to ozone for 7, 15, 30, 60 and 90 days. There is a significant increase at 60 and 90 days of treatment. (B) The results show an increase in the protein expression following 60 and 90-day exposure to ozone (*p < 0.05) compared with the control groups. GAPDH staining was used as an internal control. (C) Effects of ozone treatments on GRP78 immunoreactivity in the dentate gyrus of rat hippocampus. Light photomicrographs show immunoreactivity in GRP78 of cells from hippocampus of rats exposed to ozone. The control group showed low immunoreactivity. We observed an increase of immunoreactivity for GRP78 at 60 and 90 days of exposure to ozone. Arrows show GRP78 immunoreactivity 100×. The square indicates the area that is shown as 100× magnification.

Fig 5: NaBut modulates the repair proteins in transformed cells. (a) Immunoblotting with antibodies to repair proteins using normal (MEF, NIH3T3) and transformed (E1A+Ras) cells untreated (-) or treated with 4 mM NaBut for 48 h. Immunoblotting with a-tubulin antibodies were used as a loading control. I—hyperphosphorylated Mre11, II—hypophosphorylated Mre11, III—truncated Mre11. (b) The bar plots represent band densities, evaluated using Image J © soft, scaled to the loading control, and normalized to the band density in untreated MEF cells (folds). For Ku80 and Mre11 proteins, the relevant plots display the aggregated intensity of all detected bands at each specific point, both for untreated and NaBut-treated cells. (c) Immunoblotting with antibodies to Mre11 using E1A+Ras cells untreated (-) or treated with 2.5 µM wortmannin (WM), 20 µM LY294002 (LY), or 10 mM NaBut for 8 h. GAPDH expression was used as a loading control. (d) Immunoblotting with antibodies to Mre11 using E1A+Ras cells untreated (-) or treated with NaBut for 24–48 h. Lanes 1–3 represent total cell extracts (total CE), and lanes 4–6 represent extracts co-precipitated with Rad50 antibodies.