Journal: Virus Research

Article Title: Inhibition of EV71 replication by an interferon-stimulated gene product L3HYPDH

doi: 10.1016/j.virusres.2024.199336

Figure Lengend Snippet: Anti-EV71 activity of the endogenous L3HYPDH. (A) RT-qPCR assay of the endogenous L3HYPDH mRNA level in different cell lines, which was normalized to GAPDH mRNA level. (B) RT-qPCR assay of L3HYPDH expression with IFN-α2b (1000 IU/ml) treatment for indicated time in different cell lines. L3HYPDH mRNA level was normalized to that of GAPDH. The relative mRNA level from untreated cells (marked as 0 h) was set as 1. (C) Western blot of knockdown efficiency of shRNA-L3HYPDH. 293A-SCARB2 cells were transfected with pcDNA4-L3HYPDH together with shRNA-expressing plasmid or control plasmid (Ctrl) at a ratio of 1:3. The GFP-positive cells were sorted and then infected with EV71, followed by RT-qPCR analyses of L3HYPDH mRNA (D) and EV71 2C mRNA (E) after depression of L3HYPDH expression. RT-qPCR analyses of the L3HYPDH (F) and EV71-2C (G) mRNA in 293A-SCARB2 upon depression of L3HYPDH expression by shRNA in the presence of IFN-α2b. 293A-SCARB2 cells were seeded into a 10 cm dish and then transfected with 5 μg of pSUPER-GFP-shRNA-L3HYPDH or pSUPER-GFP. After incubation for 24 h, the GFP-positive cells were isolated using FACS and divided into two parts, one treated with 1000 IU/ml of IFN-α2b, the other mock-treated with water as control, followed by EV71-MZ infection (MOI, 0.1). Eighteen hours post-infection, total RNAs were isolated and used for RT-qPCR measurement. The target mRNA level was normalized to that of GAPDH, and the relative value from the control cells was set as 1. The results are represented as mean ± SD of three independent experiments. *, P < 0.05.

Article Snippet: Cells were treated with 1000 IU/ml of recombinant human IFN-α2b (Prospec) at different time, and then total RNAs were isolated and used to measure specific mRNA abundance by RT-qPCR.

Techniques: Activity Assay, Quantitative RT-PCR, Expressing, Western Blot, Knockdown, shRNA, Transfection, Plasmid Preparation, Control, Infection, Incubation, Isolation

Journal: The Journal of Clinical Endocrinology and Metabolism

Article Title: IFN-α Neutralizing Antibodies Distinguish LADA From Early-onset Type 1 Diabetes

doi: 10.1210/clinem/dgaf001

Figure Lengend Snippet: Correlation between AAb-IFN-α levels and GADA. Correlation between levels of AAb-IFN-α isoform 2b and GAD65 antibodies levels in individuals with LADA who tested positive for AAb-IFN-α2. Spearman's test was fitted to assess the monotonic covariation of measurements. Abbreviations: AAb-IFN-α, autoantibodies against interferon-α; GADA, glutamic acid decarboxylase antibodies; LADA, latent autoimmune diabetes in adults.

Article Snippet: The antibodies directed against the following 3 isoforms of IFN-α were evaluated in sera of study participants: IFN-α1 (InvivoGen; catalog no. rcyc-hifna1), IFN-α2b (InvivoGen; catalog no. rcyc-hifna2b), IFN-α14 (PBL Assay Science, Piscataway, NJ, USA; catalog no. 11145).

Techniques: