Horseradish peroxidase conjugated igg secondary antibody

The following reagents were purchased: LPS, from Sigma-Aldrich (St. Louis, MO, USA); Cap (purity ≥ 98%), from the National Institutes for Food and Drug Control (Beijing, China); adenovirus pAD/14-3-3γ-shRNA, from Gene Chem Co., Ltd (Shanghai, China); bafilomycin A1 (BafA1) and compound C, from Sigma-Aldrich (St. Louis, MO, USA); antibodies against 14-3-3γ, P62, NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 8 (NDUFB8) and cytochrome b-c1 complex subunit 2 (UQCRC2), from Abcam (Cambridge, UK). Anti-LC3, -AMPKα, -phospho-AMPK (phosphorylation at Ser172), -mTOR, -phospho-mTOR (phosphorylation at Ser2448), -ULK1, -phospho-ULK1 (phosphorylation at Ser757) antibodies, from Cell Signaling Technology (Beverly, MA, USA), and horseradish peroxidase-conjugated IgG secondary antibody, from Zsbio (Beijing, China).

Cap (purity ≥ 98%) was purchased from the National Institutes for Food and Drug Control (Beijing, China). Adenovirus pAD/14-3-3η-shRNA was obtained from GeneChem Co., Ltd (Shanghai, China). Antibodies directed against 14-3-3η, cytochrome c (cyt C), cleaved caspase-3, Cox4, and β-actin were obtained from Cell Signaling Technology (Beverly, MA, USA). Antibodies against NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 8 (NDUFB8) and cytochrome b-c1 complex subunit 2 (UQCRC2) were obtained from Abcam (Cambridge, UK). Horseradish peroxidase-conjugated IgG secondary antibody was purchased from Zsbio (Beijing, China).

Adenovirus pAD/Flag-AMPKα1 was purchased from GeneChem Co., Ltd. (Shanghai, China). Rg1 (purity ≥ 98%) was purchased from Solarbio (cat. no. SG8330, Shanghai, China). Compound C was acquired from Selleck (Houston, USA). Antibodies directed against AMPKα1, cytochrome b-c1 complex subunit 2 (UQCRC2), NADH dehydrogenase (ubiquinone) 1 beta subcomplex subunit 8 (NDUFB8), and Cyt C were obtained from Abcam (Cambridge, UK). Antibodies directed against Bax, Bcl-2, cleaved caspase 3, cytochrome c oxidase subunit IV (COXIV), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were obtained from Cell Signaling Technology (Boston, USA). A horseradish peroxidase-conjugated IgG secondary antibody was purchased from Zsbio (Beijing, China).
The normal rat kidney epithelial cell line (NRK-52E) was acquired from the Chinese infrastructure of cell line resources (Shanghai, China). Adult male C57BL/6 mice (8 weeks old, weighing about 20 g) were furnished by the animal center of Nanchang University (Nanchang, China). All experimental procedures followed the National Institutes of Health (NIH) Guidelines for the Care and Use of Laboratory Animals (NIH Publication No. 85-23, revised 1996) and were approved by the Ethics Committee of Nanchang University (no. 2021-0026).