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. 2021 Sep 13;12:691908. doi: 10.3389/fimmu.2021.691908

Figure 5.

Figure 5

MNSFβ promoted the expression and secretion of TNFα by binding to RC3H1 in human macrophages. (A) MNSFβ protein expression level in human Thp1-derived Mϕ as detected by Western blotting analysis (normalized to the expression of β-actin). Left panel: representative images of Western blotting assay. Right panel: the relative density of MNSFβ/β-actin. (B) MNSFβ and TNFα mRNA expression levels in Thp1-derived Mϕ as detected by RT-PCR analysis (normalized to the mRNA expression of Gapdh). (C) Invasive activity of HTR8/SVneo cells detected by transwell assay. Upper: representative images of the Transwell assay; lower: quantification of the Transwell assay by the invasion index [invasion index = X (invaded cells number)/NC (invaded cells number)]. (D) Interaction between MNSFβ and RC3H1 as validated by co-IP assay. (E) TNFα mRNA expression level in Thp1-derived Mϕ detected by RT-PCR analysis (normalized to the mRNA expression of Gapdh). (NC: Thp1-derived Mϕ transfected with NC-siRNA or HTR8/SVneo cells treated with the CM of NC Mϕ; siMNSFβ: Thp1-derived Mϕ transfected with MNSFβ-specific siRNA or HTR8/SVneo cells treated with the CM of siMNSFβ Mϕ; siMNSFβ + TNFα: HTR8/SVneo cells treated with the CM of siMNSFβ Mϕ plus TNFα; siRC3H1: Thp1-derived Mϕ transfected with RC3H1 specific siRNA; siMNSFβ + siRC3H1: Thp1-derived Mϕ transfected with MNSFβ-specific siRNA and RC3H1-specific siRNA; ns p > 0.05, *p < 0.05; ***p < 0.001; ****p< 0.0001).