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. 2022 Mar 17;36(7-9):550–566. doi: 10.1089/ars.2021.0047

FIG. 2.

FIG. 2.

NOXA1 expression is correlated with renal ROS levels in kidneys from wild-type mice treated with Ang II. (A) Real-time PCR analysis of Noxa1 mRNA expression in kidneys from male (M) and female (F) mice treated with vehicle or Ang II for 14 days. Data are mean ± SEM of mRNA expression fold change relative to vehicle-treated control adjusted for 18s RNA levels. (B) Representative immunofluorescence images and quantification of NOXA1 (red) levels in the AQP2-stained CD epithelial cells (green) in the kidney sections of male and female mice treated with vehicle or Ang II for 14 days. High magnification insets (yellow rectangle) show NOXA1 expression in CD epithelial cell. Scale is 100 μm. Data are fluorescence integrated density (mean ± SEM). (C) Real-time PCR analysis of Nox1 mRNA expression in kidneys from male wild-type and Noxa1−/− and female wild-type mice treated with vehicle or Ang II for 14 days. Data are mean ± SEM of mRNA expression fold change relative to vehicle-treated control adjusted for 18s RNA levels. (D) Representative immunofluorescence images and quantification of NOX1 (red) levels in the AQP2-positive CD epithelial cells (green) in the kidney sections of male and female wild-type and male Noxa1−/− mice treated with vehicle or Ang II for 14 days. High magnification insets (yellow rectangle) show NOX1 expression in CD epithelial cell. Scale is 100 μm. Data are fluorescence integrated density (mean ± SEM). (E) Western blot analysis and quantification of NOX1 protein expression in whole kidney lysates from mice treated with vehicle or Ang II for 14 days. Data are mean ± SEM of protein levels fold change relative to vehicle-treated control adjusted for TUBB levels. (F) ROS levels were determined by DHE fluorescence in the coronal renal sections of male and female wild-type and male Noxa1−/− mice treated with vehicle or Ang II for 14 days. Data are DHE fluorescence integrated density (mean ± SEM). (G) Superoxide levels in kidney samples from mice treated with vehicle or Ang II for 14 days were determined by 2-OH-ethidium HPLC. Data were normalized to tissue protein concentration (mean ± SEM). DHE, dihydroethidium; PCR, polymerase chain reaction; ROS, reactive oxygen species; TUBB, β-tubulin.