Journal: Biochemistry and Biophysics Reports

Article Title: MicroRNA-424 regulates the expression of CX3CL1 (fractalkine) in human microvascular endothelial cells during Rickettsia rickettsii infection

doi: 10.1016/j.bbrep.2020.100897

Figure Lengend Snippet: miR-424-5p (miR-424) expression in R. rickettsii infected endothelial cells. (A) Schematic depicting the binding site of miR-424 at the 3′UTR region of Cx3cl1 mRNA, suggesting Cx3cl1 as a potential target for miR-424. (B) Confluent HMECs were infected with R. rickettsii for 3 h or 24 h and processed using a Tri-Reagent protocol for the preparation of RNA. miR-424 expression was measured by RT-qPCR using a miR-specific Taqman assay. The error bars represent mean ± standard error from three separate experiments. The asterisks indicate significant change (p

Article Snippet: At different times post-infection, culture medium was removed by gentle aspiration and the cells were lysed directly in TRI reagent® (Molecular Research Center) for RNA isolation [ ].

Techniques: Expressing, Infection, Binding Assay, Quantitative RT-PCR, TaqMan Assay

Journal: Biochemistry and Biophysics Reports

Article Title: MicroRNA-424 regulates the expression of CX3CL1 (fractalkine) in human microvascular endothelial cells during Rickettsia rickettsii infection

doi: 10.1016/j.bbrep.2020.100897

Figure Lengend Snippet: Regulation of Cx3cl1 mRNA expression by miR-424-5p (miR-424). (A) Endothelial cells were transfected with miR-424 mimic (1nM for 24 h) prior to infection with R. rickettsii for 24 h. Cells were then lysed in Tri-reagent for the isolation of RNA and determination of miR-424 expression by a Taqman assay. (B) Cx3cl1 mRNA expression was measured by RT-qPCR using a specific primer pair. (C) CX3CL1 protein expression was measured by western blot using an antibody against CX3CL1. The asterisks indicate statistically significant change (p

Article Snippet: At different times post-infection, culture medium was removed by gentle aspiration and the cells were lysed directly in TRI reagent® (Molecular Research Center) for RNA isolation [ ].

Techniques: Expressing, Transfection, Infection, Isolation, TaqMan Assay, Quantitative RT-PCR, Western Blot

Journal: Biochemistry and Biophysics Reports

Article Title: MicroRNA-424 regulates the expression of CX3CL1 (fractalkine) in human microvascular endothelial cells during Rickettsia rickettsii infection

doi: 10.1016/j.bbrep.2020.100897

Figure Lengend Snippet: CX3CL1 expression in R. rickettsii infected endothelial cells. (A) Confluent HMECs were infected with R. rickettsii for 3 h or 24 h. At each time point post-infection, cells were lysed in Tri-Reagent for the extraction of total RNA, which was subjected to the analysis of Cx3cl1 mRNA expression by RT-qPCR using a gene-specific primer pair. (B) Total protein lysates from R. rickettsii -infected HMECs were prepared in RIPA buffer containing protease inhibitor cocktail and processed for Western blotting using an antibody against CX3CL1 to measure the levels of cellular protein expression. α-tubulin was used as a protein loading control. (C) Quantitation of CX3CL1 protein expression from three independent experiments is presented as the mean ± standard error. The asterisks indicate significant change (p

Article Snippet: At different times post-infection, culture medium was removed by gentle aspiration and the cells were lysed directly in TRI reagent® (Molecular Research Center) for RNA isolation [ ].

Techniques: Expressing, Infection, Quantitative RT-PCR, Protease Inhibitor, Western Blot, Quantitation Assay