Extended Data Fig. 1: ABHD5 is required for HDAC4 proteolysis.
From: The lipid-droplet-associated protein ABHD5 protects the heart through proteolysis of HDAC4
a, Immunoblotting using an antibody directed against endogenous ABHD5 in cardiac and cellular COS as well as HEK293 extracts. GAPDH was used as loading control. b, Cytosolic and nuclear fractions from adult rat ventricular myocytes were used for immunoblotting using an antibody directed against ABHD5, GAPDH and histone H3. a,b, Representative results of two independent experiments. c, Left, Immunoblotting analysis of cardiac extracts of control (Ctrl)- or isoproterenol (ISO)-treated mice using antibodies directed against HDAC4, ABHD5 and GAPDH as loading control. Middle left, Densitometric analysis of HDAC4-NT/FL ratio shown as fold change versus control. Middle right and right, Quantification of cardiac ABHD5 protein normalized to GAPDH and Abhd5 RNA levels normalized to 18S in unstressed mice and ISO-treated mice. Statistical analysis: Values are presented as mean ± s.e.m., n = 3 (n represents mice per indicated group); by unpaired two tailed t-test, P<0.05 considered as significant. d, Left, Immunoblotting analysis of cardiac extracts of control (Ctrl) or CL 316,243 (CL)-treated mice using antibodies directed against HDAC4, ABHD5 and GAPDH as loading control. Middle left, Densitometric analysis of HDAC4-NT/FL ratio shown as fold change versus control. Middle and Middle Right, Quantification of cardiac ABHD5 protein normalized to GAPDH and Abhd5 RNA levels normalized to 18S in unstressed mice and CL-treated mice. Right, Quantification of Ucp1 mRNA normalized to 18S in brown adipose tissue after treatment with CL as a marker of CL effective administration. Statistical analysis: Values are presented as mean ± s.e.m., n = 3 (n represents mice per indicated group); by unpaired two tailed t-test, P < 0.05 considered as significant.
