Anti tfeb antibody

Staining for cell-surface markers was performed with the following mAbs and appropriate isotype controls (all BD Biosciences) for 30 min at 4°C; anti-CD86, anti-CD4, anti-CD8, anti-CD11c, anti-CD11b, anti-CD19. Anti I-A^b mAb was obtained from Affymetrix. H-2K^b – associated with the OVA SIINFEKL peptide was detected with the mAb 25.D1. TFEB was detected in fixed and permeablized cells with anti-TFEB antibody from MyBioSource, Data was collected on BD Accuri flow cytometer system (BD Biosciences) and analyzed with FlowJo software.

De-paraffinized tissue sections were antigen-unmasked and wheat germ agglutinin staining and Masson’s trichrome staining were performed as previously described (21 (link),24 (link)). For GFP-LC3 dot visualization in Tg-GFP-LC3 mice, myocardial samples were embedded in Tissue-Tek OCT compound (Sakura Finetechnical Co. Ltd) and stored at −80°C. The samples were sectioned at 5–7 μm thickness with a cryostat (CM3050 S, Leica). GFP-LC3 dots were observed under a fluorescence microscope as previously described (17 (link),21 (link)).
For immunofluorescent staining, fixed cardiomyocytes were incubated overnight with anti-TFEB antibody (MyBioSource) and then with Alexa Fluor 568 dye-conjugated secondary antibody (Life Technologies). Samples were washed and mounted on glass slides with DAPI (Vectashield; Vector Laboratories).