Plin1

The isolated macrophages were lysed in ice‐cold RIPA buffer. The supernatant was collected and stored at −80°C. Proteins were separated on 10%–15% sodium dodecyl sulphate‐polyacrylamide gel by electrophoresis and then transferred to nitrocellulose membrane (Pall BioTrace NT). After blocking with 3% bovine serum albumin containing 0.1% Tween 20 for 1 h at room temperature, target proteins were detected by incubating with primary antibodies at 4°C overnight. The following primary antibodies were used: from Cell Signalling Technology (CST), p‐p65 (1:500; #3033), p65 (1:1000; #8242), LC3BI/II (1:500; #2775), SQSTM1/p62 (1:1000; #5114), Becline‐1 (1:1000; #3738), p38 MAPK (1:1000; #9212), p‐p38 MAPK (1:2000; #4631), ATGL (1:1000; #2439), HSL (1:1000; #4107), p‐HSL (1:500; #4139), p‐PKCδ (1:500; #2055), PKCδ (1:1000; #2058), and GAPDH (1:5000; #5174); from Vala Sciences, PLIN1 (1:5000; #4854) and p‐PLIN1 (1:5000; #4856); and from Santa Cruz, P38IP (1:100; SC‐374665). After overnight incubation, membranes were probed with horseradish peroxidase‐conjugated secondary antibody (CST, #7074) for 1 h at room temperature. Signals were then detected with chemiluminescence reagent (LumiGlo; CST, #7003). Images were taken using ChemiDoc XRS imager (Bio‐Rad). Band intensity was determined by densitometry analysis using Quantity One® software and normalised using GAPDH as the loading control.

Total proteins from MCF-7 and MDA-MB-231 cell lysates were extracted by resuspending the cell pellets in RIPA buffer (150 mM NaCl, 50 mM Tris (pH 7.4) and 1% Triton X-100). Approximately 55 μg of total protein per sample was separated by SDS-PAGE and then transferred onto nitrocellulose membranes. Western blot analyses were performed with polyclonal antibodies against PLIN1 (Santa Cruz Biotechnology, USA), with a monoclonal β-actin antibody as a control (Sigma, USA).