Fig 1: NLRP3 deficiency reduces crosstalk between VSMCs and macrophages. A, mRNA expression levels of IL‐1α, IL‐1β, IL‐6, Tnf‐α, Ccl2, Cxcl12, and Cx3cl1 in primary VSMCs from NLRP3 fl/fl and NLRP3 SMCKO mice after 24 h starvation and 48 h 20% CKD serum stimulation. B, Bone marrow–derived monocytes from WT mouse were seeded on Transwell dishes to perform migration assays. Primary VSMCs from NLRP3 fl/fl and NLRP3 SMCKO mice after 24 h starvation and 48 h 20% CKD serum stimulation were used as a chemoattractant. Cells migrating through the membrane were fixed after 24 h (left) and the cell number per HPF was counted (right). C, Primary VSMCs from NLRP3 fl/fl or NLRP3 SMCKO mice after 24 h starvation and 48 h 20% CKD serum stimulation were harvested and then injected into WT mice intraperitoneally. The peritoneal lavage fluid was collected for flow cytometry after 12 h. D, The proportion (left) and estimated counts (right) of macrophages (F4/80+, MHC II+), B cells (F4/80−, MHC II+), and T cells/neutrophils (F4/80− and MHC II−) were calculated. E, The activation of macrophages was assayed by expressions of MHC II, CD40, and CD80. Quantification is shown on the right. Scale bars: 20 μm. *P<0.05; ns, not statistically significant; data were analyzed by the unpaired t test. CCL2 indicates C‐C Motif Chemokine Ligand 2; CKD, chronic kidney disease; CX3CL1, C‐X3‐C Motif Chemokine Ligand 1; CXCL12, C‐X‐C Motif Chemokine Ligand 12; IL, interleukin; Mac, macrophages; mRNA, messenger RNA; NLRP3, nucleotide‐binding oligomerization domain‐like receptor protein 3; NLRP3 SMCKO, smooth‐muscle‐specific knockout nucleotide‐binding oligomerization domain‐like receptor protein 3; N/T, neutrocytes or T cells; PLF, peritoneal lavage fluid; TNF, tumor necrosis factor; VSMC, vascular smooth muscle cell; WT, wild‐type.