Western Blot Analysis of Autophagy, PCSK9, and SIRT4 Proteins

Protein lysis, sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE), nitrocellulose membrane transfer, and chemiluminescent acquisition and analysis were conducted as already described [11 (link), 50 (link)] with anti-microtubule-associated proteins 1A/1B light chain 3B (LC3B II/I, 1:2000, ab192890, Abcam, Cambridge, UK), anti-PCSK9 (1:1000, ab185194, Abcam, Cambridge, UK), anti-SIRT4 (1:1000, ab231137, Abcam, Cambridge, UK), anti-α-tubulin (1:5000, E-AB-20036, Elabscience Biotechnology Inc., Houston, TX, USA), and anti-actin (1:3000, ab179467, Abcam, Cambridge, UK) as primary antibodies.