Journal: mBio

Article Title: Interferon-Lambda Intranasal Protection and Differential Sex Pathology in a Murine Model of SARS-CoV-2 Infection

doi: 10.1128/mBio.02756-21

Figure Lengend Snippet: IFN-λ inhibited SARS-CoV-2 replication in cultured human lung cells. Human bronchial epithelial cells expressing hACE2 (HBE-hACE2) were either mock treated (black-filled circles), pretreated for 24 h with IFN-λ (pre-IFN-λ) (green open squares), or treated with IFN-λ at the same time of infection (co-IFN-λ) (green-filled circles) with SARS-CoV-2. Culture medium was harvested at 0, 24, and 48 h postinfection, and viral RNA in culture medium was measured by RT-qPCR following infection at MOI of 0.01 PFU/cell (A) or MOI of 0.1 PFU/cell (B). Mature virus released into the culture media was quantified by plaque assay following infection at MOI of 0.01 PFU/cell (C) or MOI of 0.1 PFU/cell (D). Three independent experiments were performed with technical duplicates. * , P < 0.001 (two-way ANOVA). (E) Representative Western blots are shown of proteins expressed in the HBE and HBE-hACE2 cells following treatment with increasing amounts of IFN-λ (200 to 2,000 units/ml) for 24 h. (F) RT-qPCR measurements are shown for ISG mRNAs expressed in HBE-hACE2 cells relative to untreated controls in response to IFN-λ treatment (1,000 units/ml) or relative to mock treatment in response to a SARS-CoV-2 infection for 24 h.

Article Snippet: Human bronchial epithelial (HBE) 3-KT cells were obtained from American Type Culture Collection (ATCC) and cultured in airway epithelial cell basal medium supplemented with the bronchial epithelial cell growth kit (ATCC).

Techniques: Cell Culture, Expressing, Infection, Quantitative RT-PCR, Virus, Plaque Assay, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: miR-21-mediated Radioresistance Occurs via Promoting Repair of DNA Double Strand Breaks *

doi: 10.1074/jbc.M116.772392

Figure Lengend Snippet: miR-21 modulated radioresistance in human cells. A, illustration of NanoString measurement. miRs are specifically ligated to unique tags for downstream detection (top panel). Capture and reporter probes are shown in the bottom panel. B, the relative levels of miR-21 (ratio of tumor to non-tumor in each pair of tissue samples) in 30 pairs of human lung cancer tissue and their adjacent non-tumor tissue and in 14 NSCLC cell lines compared with two normal human bronchial epithelial cell lines were measured using NanoString technology. The red line indicates the miR-21 levels in the normal controls. C, the sensitivities of two miR-21 stably overexpressed NL20 cell lines, miR-21-1 and miR-21-2) or without miR-21 (vector transfection control), to IR were measured using a clonogenic assay. The data are the mean ± S.D. and were obtained from three separate experiments. D, the NSCLC cell lines (1, A549; 2, H460; 3, H358; 4, H2792) were treated with either control RNA or anti-miR-21 RNA. At 48 h after transfection, the cells were exposed to 4 Gy and then the radiosensitivity was measured using a clonogenic assay. The data are the mean ± S.D. and were obtained from three separate experiments.

Article Snippet: To verify the effects of miR-21 on radioresistance in human cells, we initially compared the miR-21 levels in 30 pairs of human lung cancer tissue with their adjacent non-tumor tissue in wax blocks (obtained from the Department of Pathology and Laboratory Medicine at the Emory University School of Medicine) as well as in 12 human non-small cell lung cancer (NSCLC) cell lines (compared with two human bronchial epithelial cell lines, 3KT (obtained from Dr. Mina's lab ( 32 )), and NL20 (purchased from the American Type Culture Collection (ATCC)) using NanoString technology ( 33 ) ( A ).

Techniques: Stable Transfection, Plasmid Preparation, Transfection, Clonogenic Assay