Cell culture and treatments
The mouse macrophage cell line (Raw 264.7) and mouse kidney proximal
tubular cell line (TCMK1) were obtained from Shanghai Institute of
Biochemistry and Cell Biology, Chinese Academy of Sciences. Raw 264.7
(RRID: CVCL_0493) and TCMK1 (RRID: CVCL_2772) cells were cultured in
RPMI 1640 and DMEM/F12 media (both were supplemented with 10% FBS),
respectively. To investigate whether tubular cell is the essential
mediator of effect from uric acid on macrophage, we co-cultured Raw
264.7 and TCMK1 together by putting transwell inserts carrying TCMK1 to
6-well plates with Raw 264.7. Cells were exposure to uric acid at
concentration of 800 μM, as previously reported(Ren et al., 2021). Based
on different intervention purposed, before adding uric acid, cells were
treated with Itgam siRNA (Genepharma Co., Ltd., Suzhou, China), FAK
inhibitor (PF562271, Selleck Chemicals LLC, USA), and Akt inhibitor
(MK-2206 2HCl, Selleck Chemicals LLC, USA) as instructed.