Fig. 4: DPEP2 deficiency enhances inflammatory response of macrophages in vitro.

A mRNA expression levels of inflammatory cytokines (IL-1β, IL-6, and TNF-α) in WT, DPEP2-KO, and DPEP2-KO + DPEP2 re-expression (rDPEP2) iBMDMs treated with LPS treatment (1 μg/ml, 6 h). n = 3 per group. B ROS production levels in WT, DPEP2-KO, and DPEP2-KO + DPEP2 re-expression (rDPEP2) iBMDMs treated with LPS treatment (1 μg/ml, 6 h). Representative images of ROS production were in up and statistical analysis was in down. n = 3 per group. C Western blot assays showing IL-1β and MyD88 dependent changes in IκBα levels, p-p38, and p-ERK1/2 at indicated time points of treatment with LPS (1 μg/ml) in WT, DPEP2-KO, and DPEP2-KO+ rDPEP2 iBMDMs. D The complex of MyD88 and IRAK4 detected by Co-IP and Western blot analysis 30 min post treatment with LPS (1 μg/ml) in WT, DPEP2-KO, and DPEP2-KO+rDPEP2 iBMDMs. E Indicating active (dimerized) IRF3 detected by Western blot assays using NativePAGE electrophoresis 30 min post treatment with LPS (1 μg/ml) in WT, DPEP2-KO, and DPEP2-KO+rDPEP2 iBMDMs. F NF-κB expression levels in nucleus detected by nuclear protein extraction and Western blot assays 60 min post treatment with LPS (1 μg/ml) in WT, DPEP2-KO, and DPEP2-KO+ rDPEP2 iBMDMs. G The detection of NF-κB activation as an inflammatory transcription factor by ChIP-qPCR targeting Nlrp3, CCL11, and NFKBIA promoter in WT, DPEP2-KO, and DPEP2-KO+ rDPEP2 iBMDMs with LPS treatment. n = 3 per group. All data are expressed as mean ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001.