A16678

Immunoblot assays were performed according to standard protocol. Target proteins were detected with antibodies against NLRP3 (A12694, Abclonal, Wuhan, China), collagen I (A5786, Abclonal, Wuhan, China), fibronectin (A16678, Abclonal, Wuhan, China), Flag (F1804, Sigma, St Louis, USA), caspase1 (A0964, Abclonal, Wuhan, China), IL-1β (A11369, Abclonal, Wuhan, China), IL-18 (A1115, Abclonal, Wuhan, China), phos-smad3 (AP0727, Abclonal, Wuhan, China), E-cadherin (3195, CST, Danvers, USA), N-cadherin (14215, CST, Danvers, USA), FIP1 (A7138, Abclonal, Wuhan, China), β-actin (AC026, Abclonal, Wuhan, China).

MSC(AT)s were seeded in 24-well plates and incubated in a complete medium. After MSC(AT)s reached ~50% confluency, they were prepared for IF staining. HGFs were seeded in 24-well plates and divided into four groups (see Section 4.6, Cell migration assay). For cell IF staining, cells were washed with PBS three times and fixed with 4% paraformaldehyde for 15 min, after which they were treated with 0.1% Triton X-100 for 20 min, blocked by 5% normal goat serum for 30 min at RT, and incubated at 4 °C overnight with antibodies, COL1A1 (A16891, Abclonal, Wuhan, China), IL-1RA (ab124962, Abcam, Cambridge, UK), and FN (A16678, Abclonal, China). The next day, cells were rewarmed at room temperature (RT) for 30 min and incubated with secondary antibodies (ab96899, Abcam, Cambridge, UK or ZLI-0316, ZSGB-BIO, China) for 1 h. After being washed with PBS three times for 5 min each, they were stained with a fluorescent mounting medium with DAPI. Images were acquired with an Olympus microscope (Olympus Corporation, Tokyo, Japan).