Fig. 1: Degradation of PGC1α is induced by CMA under thermal stress.

a, b Immunoblotting for PGC1α protein levels in HEK293T cells cultured in 37 °C or 39 °C for 24 h (representative of n = 3 independent experiments). c Immunoblotting for PGC1α protein levels in MEF cells with/without ATG5 deletion or overexpressing VPS4A or VPS4AE228Q (dominant negative) in 39 °C for 24 h and treated with 50 μM cycloheximide (CHX) for 3 h before harvest cells (representative of n = 3 independent experiments). d Immunoblotting for PGC1α and LAMP2A protein levels in HEK293T cells with/ without LAMP2A knockdown in 39 °C for 24 h and treated with 50 μM cycloheximide (CHX) for 3 h before harvest cells (representative of n = 3 independent experiments). e Immunoblotting for PGC1α and LAMP2A protein levels in HEK293T cells over-expressing LAMP2A in 39 °C for 24 h and treated with 50 μM cycloheximide (CHX) for 3 h before harvest cells (representative of n = 3 independent experiments). f Immunoblotting for PGC1α and LAMP2A protein levels in HEK293T cells treated with QX77 in 39 °C for 24 h and treated with 50 μM cycloheximide (CHX) for 3 h before harvest cells (representative of n = 3 independent experiments). g, h Immunoblotting for PGC1α and UCP1 protein levels in primary brown adipocytes in 39 °C for 24 h (representative of n = 3 independent experiments). i Immunoblotting for PGC1α, UCP1 and LAMP2A protein levels in primary brown adipocytes overexpressing LAMP2A in 39 °C for 24 h and treated with 50 μM cycloheximide (CHX) for 3 h before harvest cells (representative of n = 3 independent experiments). j Immunoblotting for PGC1α protein levels in HEK293T cells overexpressing PGC1α wt/ mutants in 39 °C for 24 h and treated with 50 μM cycloheximide (CHX) for 3 h before harvest cells (representative of n = 3 independent experiments). k Immunoblotting for PGC1α and LAMP2A protein levels in HEK293T cells co-expressing LAMP2A and PGC1α -MUT in 39 °C for 24 h and treated with 50 μM cycloheximide (CHX) for 3 h before harvest cells (representative of n = 3 independent experiments). l Immunoblotting for PGC1α protein levels in HEK293T cells overexpressing PGC1α wt/ mutants in 39 °C for 24 h. m Endogenous Co-IP analysis of HSC70 and LAMP2A in BAT from mice housed at RT and TN (n = 3 mice for 23 °C group, n = 4 for 29 °C group). n Immunoblotting for LAMP2A protein level in BAT from mice housed at RT and TN. o Representative cell images and quantitation of CMA flux of NIH-3T3 cells expressing KFERQ-PA-mCherry1 (n = 20), Scale bar, 50 µm. Statistical significance was assessed by unpaired Student’s t-test. All results were expressed as means ± SEM and **p < 0.01, ***p < 0.005. Student’s two-tailed unpaired T-test for 2-group comparisons. Source data are provided as a Source Data file.