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Additional file 1: Fig. S1. Methyl-β-cyclodextrin efficiently clears accumulated cholesterol in the presence of U18666A and bafilomycin A1. A RPE1 cells were treated with or without different concentration of MβCD in a serum-starvation medium for 24 hours. Cells were then subjected to Western blot for LC3, SQSTM1, phospho-RPS6, RPS6, and ACTB. B Cells were either treated with U18666A (2 μg) or bafilomycin (25 nM) for 12 hours in a serum-starvation medium. Cell were further incubated for additional 24 hours or incubated for additional 24 hours with MβCD (10 μM or 20 μM) without changing the medium. Cells were then subjected to filipin staining. Scale bar, 20 μm. C, D Fluorescence intensities of filipin were measured by ImageJ Software. Filipin intensity from 50 cells were acquired for each experimental group. Bar graph represents mean ± SD (n = 3 experiments). *P < 0.05, Student's t-test. Fig. S2. Methyl-β-cyclodextrin enhances LAMP1 localization to GFP-LC3 positive structure in the presence of U18666A and bafilomycin A1. A RPE1 cells were treated with U18666A (2 μg), chloroquine (5 μM) or bafilomycin A1 (25 nM) for 12 hours in a serum-starvation medium. Cells further incubated with or without MβCD (20 μM) for additional 24 hours without changing medium. GFP-LC3 transfection transfection was carried out and after 6 hours of transfection cells were fixed by 4% paraformaldehyde and immunostained with LAMP1 antibody. Scale bar, 10 μm. B Quantification of GFP-LC3 puncta overlapping with LAPM1 was quantified from 30 transfected cells by using ImageJ software. Bar graph represents mean ± SD (n = 3 experiments). #P < 0.05, Student's t-test (test group were compared with control). *P < 0.05 Student's t-test (respective test groups were compared in the presence or absence of MβCD).

Additional file 1: Fig. S1. Catalase expression was specifically increased during sustained fasting. Fig. S2. The expression of catalase was successfully depleted in mice. Fig. S3. Sustained fasting decreased the level of free fatty acid in serum of catalase KO mice. Fig. S4. Catalase deficiency during sustained fasting did not induce inflammatory response, and lipodystrophy in adipocytes. Fig. S5. Catalase deficiency during sustained fasting did not show any morphological change in BAT. Fig S6. Catalase activity was significantly increased during sustained fasting with no change in other antioxidant enzyme. Fig. S7. Lipolysis by isoproterenol increased the peroxisomal enzyme in primary BAT of catalase KO mice. Table S1. List of antibodies used in this study. Table S2. List of primer sequence used for Q-PCR.