Fig 2: FZHY regulated the gene expression of chemokines CCL2 and CX3CL1 in primary intrahepatocyte macrophages. Results are expressed as mean ± SD (n = 3). ##p < 0.01 vs. normal group, *p < 0.05 and **p < 0.01 vs. CCl4 group.

Fig 3: iRHOM2 interacts with CX3CL1 to affect endothelial integrity in LPS-induced human pulmonary microvascular endothelial cells. (A) Analysis of endothelial barrier permeability using TEER measurement. (B) Immunofluorescence staining for ZO-1 protein expression. (magnification, x100). (C) Western blot assay was performed to analyze protein expression of ZO-1, VE-cadherin and occludin. Data are presented as the mean ± standard deviation of three independent experiments performed in triplicate. ***P<0.001 vs. control. ##P<0.01 and ###P<0.001 vs. LPS. +P<0.05, ++P<0.01 and +++P<0.01 vs. LPS + siRNA-iRHOM2 + Ov-NC. ZO-1, zonula occludens-1; iRHOM2, inactive rhomboid-like protein 2; siRNA, small interfering RNA; NC, negative control; LPS, lipopolysaccharide; VE, vascular-endothelial; TEER, transendothelial electrical resistance.

Fig 4: iRHOM2 interacts with CX3CL1 to affect apoptosis in response to LPS treatment. (A) Apoptosis levels were measured using TUNEL assay. (Magnification, x100). (B) Western blotting was performed for measurement of Bcl-2, Bax and cleaved caspase3 protein levels. Data are presented as the mean ± standard deviation of three independent experiments performed in triplicate. ***P<0.001 vs. control. ###P<0.001 vs. LPS. ++P<0.01 and +++P<0.01 vs. LPS + siRNA-iRHOM2 + Ov-NC. CX3CL1, C-X3-C motif chemokine ligand 1; iRHOM2, inactive rhomboid-like protein 2; siRNA, small interfering RNA; NC, negative control; LPS, lipopolysaccharide; Ov, overexpression.

Fig 5: CX3CL1 expression is decreased by iRHOM2 interference in LPS-treated human pulmonary microvascular endothelial cells. (A) RT-qPCR analysis of CX3CL1 expression in response to LPS stimulation. (B) Western blot analysis of CX3CL1 in response to LPS stimulation. (C) Reverse transcription-quantitative PCR analysis was performed to measure mRNA expression of CX3CL1. (D) Western blot assay was performed to measure protein levels of CX3CL1. (E) Immunoprecipitation assay showed the existence of CX3CL1 in anti-iRHOM2. (F) Immunoprecipitation assay showed the existence of iRHOM2 in anti-CX3CL1. association between iRHOM2 and CX3CL1. Data are presented as the mean ± standard deviation of three independent experiments performed in triplicate. **P<0.01, ***P<0.001 vs. control. ###P<0.001 vs. LPS + siRNA-NC. RT-qPCR, reverse transcription-quantitative polymerase chain reaction; CX3CL1, C-X3-C motif chemokine ligand 1; iRHOM2, inactive rhomboid-like protein 2; siRNA, small interfering RNA; NC, negative control; LPS, lipopolysaccharide.