Anti lamin b 12987 1 ap

2% SDS buffer with 1% protease inhibitor cocktail (P8340, Sigma-Aldrich) was used to lyse renal cortex and outer medullary tissue. The protein concentration was determined using the Pierce BCA Protein Assay Kit from Thermo Scientific. The primary antibodies used for immunoblotting included: anti-Fibronectin (ab2413), anti-α-Smooth Muscle Actin (ab5694), anti-IL-6 (ab229381) and anti-TNF-α (ab183218) from Abcam; anti-STAT1(D1K9Y) (14994), anti-phospho-STAT1(Y701) (9167), anti-HMGB1(D3E5) (6893), anti-NF-κB p65 (8242), anti-phospho-NF-κB p65 (3033), anti-GAPDH (5174), anti-Vimentin (5741), anti- Histone H3 (1B1B2) (14269) and anti-IL-1β (12242) from Cell Signaling Technology; anti-Collagen type I (AF7001) from Affinity. anti- Lamin B (12987-1-AP) from Proteintech. All secondary antibodies for immunoblot analysis were from Thermo Fisher Scientific (MA, USA). The samples were resolved on SDS-polyacrylamide gel, and then transferred for immunoblotting. The PVDF membrane is incubated with 5% BSA for 1 h. The membranes were then incubated overnight with primary antibodies according to the ratios in the instructions. After rewarming, the secondary antibody was incubated for 1 h. Finally, the chemiluminescence imaging system was used for luminescence imaging. For densitometry, the protein bands were analyzed with the NIH ImageJ software.

We used the following commercially available antibodies: The anti-AdipoR1 (ab126611), anti-FN (ab2413), anti-Collagen I (ab34710), anti-AMPKα1 + AMPKα2 (ab131512), anti-AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172) (ab23875), anti-RAB7(ab50533) and anti-ADRP (ab52356) antibodies were from Abcam; the anti-phospho-AMPKα(Thr172) (2535), anti-mTOR(2972), anti-phospho-mTOR(Ser2448)(5536), anti-ULK1 (8054), anti-phospho-ULK1 (Ser555) (5869), anti-SQSTM1/p62(5114) and anti-LC3B (3868) antibodies were from Cell Signaling Technology; and the anti-SQSTM1/P62 (18420-1-AP), anti-LC3B (18725-1-AP), anti-Beclin1 (11306-1-AP), anti-ADRP (15294-1-AP), anti-ATG5 (10181-2-AP), anti-TFEB (13372-1-AP), anti-β-actin (60008-1-Ig), anti-GAPDH (60004-1-Ig) and anti-Lamin B(12987-1-AP) antibodies were from Proteintech. The anti-SREBP-1c(sc-13551) was from Santa Cruz Biotechnology. AdipoRon and CQ was obtained from Selleckchem. AICAR was provided by Apexbio Technology, SBI-0206965 was purchased from Sigma-Aldrich.