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. 2021 Jul 8;6(13):e147505. doi: 10.1172/jci.insight.147505

Figure 5. Deletion of ZC3H12A gene elevates inflammatory mediator expression in the HK-2 cell line.

Figure 5

(A) Experimental strategy to delete ZC3H12A gene by CRISPR-Cas9 in the HK-2 cell line. (B) HK-2 and HK-2ΔZC3H12A cells were lysed and evaluated for Regnase-1 by Western blot. Protein relative abundance and representative image are shown. (C, E, and G) HK-2 and HK-2ΔZC3H12A cells were stimulated with IL-17 and/or TNF-α and mRNA expression of IL6, LCN2, and NFKBIZ was measured by qPCR (8 hours after stimulation), normalized to GAPDH. (D and F) Protein level of IL-6 (8 hours after stimulation) and LCN2 (24 hours after stimulation) in culture supernatant were measured by ELISA. (H) HK-2 and HK-2ΔZC3H12A cells were stimulated with IL-17 and/or TNF-α for 8 hours, and cell lysates were evaluated for IκBξ by Western blot. Protein relative abundance and representative image are shown. Data pooled from at least 3 independent experiments. Statistical analysis by 2-tailed unpaired t test (B) or 2-way ANOVA (C–H). *P < 0.05; **P < 0.01; ***P < 0.001.