Journal: Neurogenetics

Article Title: Loss of mitochondrial ClpP, Lonp1, and Tfam triggers transcriptional induction of Rnf213 , a susceptibility factor for moyamoya disease

doi: 10.1007/s10048-020-00609-2

Figure Lengend Snippet: RT-qPCR analyses of wild-type MEF, human neuroblastoma SH-SY5Y cells, and human umbilical vein endothelial cells (HUVEC) for the expression of Rnf213 after exposure to different stress situations. Rnf213 transcript in a MEF, b SH-SY5Y, and c HUVEC cells after serum starvation (DMEM, Dulbecco’s modified Eagle medium; FCS, fetal calf serum; HBSS, Hank’s balanced salt solution; CTRL, untreated control) for indicated times. Rnf213 transcript in d MEF, e SH-SY5Y, and f HUVEC cells is quantified after application of the pathogenic dsRNA analog Poly(I:C) for 16. Rnf213 transcript in g MEF, h SH-SY5Y, and i HUVEC cells is quantified after incubation with the bacterial cell wall component Lipopolysaccharide (LPS) for 24 h. R nf213 transcript in j MEF, k SH-SY5Y, and l HUVEC cells is quantified after incubation with murine or human interferon gamma (IFNG). The Y -axis of each plot shows the ratio of a transcript of interest versus mouse Tbp or human HPRT1 as loading control. The bar graphs show mean and standard error of the mean (SEM), illustrating the significances with asterisks (Trend T 0.05 < p < 0.1; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). m Quantitative immunoblot for RNF213 protein expression in untreated WT and ClpP −/− MEF cells, and after incubation with Poly(I:C) at 1 μg/ml for 16 h. HSP90 served as loading control

Article Snippet: Recombinant mouse IFNG (Merck Millipore, IF005) or human IFNG (Preprotech, 300-02) was applied to MEF, SH-SY5Y and HUVEC cells ( n = 3–6, each) at 50 ng/ml for 24 h. Cells were collected for RNA extraction.

Techniques: Quantitative RT-PCR, Expressing, Modification, Control, Incubation, Western Blot