Ab172907

Immunofluorescent staining was performed using primary antibodies targeting EGR1 (1:100 dilution; ThermoFisher, PA5-83115), MFGE8 (1:100 dilution; ThermoFisher, PA5-82036), MMP1 (1:100 dilution; Abcam, ab52631), α-smooth muscle actin (1:200 dilution; Abcam, ab5694), YAP (1:100 dilution; CellSignaling, 14074 S), and Perilipin-1 (1:100 dilution; Abcam, ab172907). The percentage of fluorescent area was quantified using custom a MATLAB image processing code written by the authors and previously published^{26 (link)}. All histology and immunofluorescent images shown are representative images of multiple experiments.

Cells or frozen sections of OC tissues were fixed with 4% paraformaldehyde for fifteen minutes, permeabilized with 0.3% Triton X-100 for 15 min, and blocked with 5% goat serum (Life Technologies, USA) for 1 h at room temperature. Then, sections were incubated with primary antibodies for 12 h at 4 °C. Secondary antibodies were added, and staining with DAPI (Life Technologies, USA) was then performed. Stained sections were imaged using a Leica SP5 confocal fluorescence microscope. Primary antibodies specific for the following proteins were used in the IF experiments: STC1 (sc-293435, Santa Cruz Biotechnology, 1:100 dilution, Dallas, Texas, USA), FOXC2 (23,066–1-AP, Proteintech, 1:100 dilution, Wuhan, Hubei, China), ITGB6 (19,695–1-AP, Proteintech, 1:100 dilution, Wuhan, Hubei, China), UCP1 (AF8292, Beyotime, 1:100 dilution, Shanghai, China), TOM20 (11,802–1-AP, Proteintech, 1:100 dilution, Wuhan, Hubei, China), perilipin1 (ab172907, Abcam, 1:100 dilution, Cambridge, UK). All antibodies are listed in Supplementary Table S1 (The primary antibodies). The secondary antibodies were FITC- or Texas red-conjugated donkey F(ab)2 fragments specific for mouse IgG or rabbit IgG (Jackson ImmunoResearch Laboratories).

The antibody against Senp7 (LS-C98791) was purchased from LSBio, the antibody against SUMO2/3 (ab81371) and Plin1 (ab172907) were purchased from ABCam, the antibody against Plin2 (15294-1-ap) was purchased from Proteintech, the antibody against Plin3 (NB110-40764SS) was purchased from NOVUS, the antibody against Plin4 (ABS526), PLIN4 (HPA044682), and Flag (F1804) were purchased from Sigma, the antibody against Myc (#2276) and HA (#3724) were purchased from CST, and the antibody against α-tubulin (BS1699) was purchased from Bioworld Technology, Inc. All the primary antibodies were diluted in a ratio of 1:1000. Secondary antibodies, anti-mouse IgG and anti-rabbit IgG, were purchased from Sigma-Aldrich. All the secondary antibodies were diluted in a ratio of 1:10,000.