NLRP3 Inflammasome Simultaneously Involved in Autophagy and Phagocytosis of THP-1 Cells to Clear Aged Erythrocytes

Figure 1

Results of CRISPR/Cas9 editing for NLRP3 in THP-1 cells and the stable transfectant THP-1 cell line. (a) Use of three programs to edit exons 9 (ID3), 7 (ID4), and 5 (ID5) of the NLRP3 gene in THP-1 cells. The shNC indicated nontargeting control. The expression of NLRP3 was determined via QRT-PCR. All three schemes can significantly downregulate the NLRP3 gene expression, and ID3 obtained the optimal regulation results. ID3 THP-1 was used in the follow-up study. (b)–(d) Microscopic examination results (100×) of THP-1 cells stably transfected with shNC and ID3 vector. Cells demonstrated normal morphology and ability to proliferate and generate. (c) NLRP3 expression in the stable transfectant ID3 THP-1 cell line tested via QRT-PCR. The expression of NLRP3 in ID3 THP-1 was still stable and low after more than three passages. (e) Evaluation of the regulatory effect of three gene-editing schemes on NLRP3 via immunoblot. ID3 showed optimal ability to downregulate the NLRP3 expression. (f) NLRP3 expression in the stable transfectant ID3 THP-1 cell line tested using immunoblot. The result was similar to that obtained via QRT-PCR. , ns: not significant.

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