Mouse anti human vimentin

The cells were lysed within RIPA buffer that contained 1% PMSF, and the proteins were loaded onto SDS‐PAGE microgels and transferred to PVDF membranes. Then 50 g/L skim milk powder was managed for blockage, and the primary antibodies diluted in 50 g/L BSA were added, including rabbit anti‐human c‐met (1: 100, Abjent, USA), mouse anti‐human E‐cadherin (1:3000, Abcam, Cambridge, MA, USA), mouse anti‐human N‐cadherin (1:8000, Abcam, Cambridge, MA), mouse anti‐human vimentin (1:1000, Cell Signaling Technology, Danvers, MA, USA) and mouse anti‐rat α‐SMA monoclonal antibody (1:300, Boster, Wuhan, China). They were placed at 4°C for overnight before incubating the blots with HRP‐conjugated goat anti‐mouse IgG (1:500, ZSGB‐BIO, Beijing, China). Darkroom visualization was achieved using ECL substrate visualization signals (Millipore, Billerica, MA, USA), and GAPDH was used as normalized endogenous white matter.

Frozen endometriotic lesions were embedded in optimal cutting temperature compound and sectioned at a 5 μm thickness. Then, the sections were washed with PBS, permeabilized with 0.2% Triton X-100, washed with PBS for 10 minutes, and blocked with 10% FBS for 1 hour. The sections were then incubated with primary antibodies overnight at 4°C, washed with PBST, and then, where required, stained with Alexa Fluor–conjugated secondary antibodies for 1 hour at room temperature. The following primary antibodies were used: Goat Anti-human IL-33 (1:40, R&D Systems, AF3625), Mouse Anti-human Vimentin (1:400, Cell Signaling Technology, clone D21H3), Mouse Anti-Pan cytokeratin (1:40, Cell Signaling Technology, clone AE1/AE3), Rabbit Anti-goat Alexa Fluor 647 (1:200; Invitrogen, A27018). Immunostained tissue sections were imaged using the Leica TCS SP8 confocal microscope. Images were taken in a Z-stack (1.20 μm; Z dimension) and tile scan (12 tiles, 580 μm × 435 μm each tile; x and y dimensions) acquisition mode. Two to 3 fields per preparation were imaged using HC PL APO CS2 ×63/1.40 oil objective and LAS-X Software (Leica Microsystems).