Journal: Molecular and Cellular Biology

Article Title: In vivo evidence of interaction between interferon-stimulated gene factors and the interferon-stimulated response element.

doi: 10.1128/mcb.9.8.3533

Figure Lengend Snippet: FIG. 2. Comparison of ISG15 constitutive and IFN-a-induced DNase-hypersensitive sites in WI38VA and Daudi cells at different times after IFN-a treatment. (A) WI38VA cells were treated with 500 U of IFN-a per ml for the time shown. Nuclei were isolated and treated with the indicated concentrations of DNase, and then DNA was extracted, digested with TaqI, electrophoresed, blotted, and probed with a radiolabeled ISG15 EcoRI-Taql fragment (+510 to +850). Fragment sizes (base pairs) were calculated with the equa- tion obtained from linear regression analysis of the marker mobili- ties (R2 > 0.99), and the centers of sites were then estimated on the basis of known location of the probe within the gene. Resolution in the size range of the promoter was very good, and the centers of the DNase-hypersensitive sites were reproducibly mapped to within +10 base pairs. The uppermost site is the full-length restriction fragment detected by the probe. The markers (M) used were 1-kilobase and 123-base-pair ladders (Bethesda Research Laborato- ries) labelled with 32P and mixed with 20 ,ug of sheared salmon sperm DNA prior to electrophoresis. (B) Daudi cells were treated with 250 U of IFN-a per ml for the time shown. The analysis was performed as described for panel A. (C) The results illustrated by panels A and B are depicted schematically. Restriction sites and DNase-hyper- sensitive sites I through IV are shown above the horizontal line. The induced hypersensitive site is marked by the heavy vertical line. The positions given for the hypersensitive sites represent the centers of the analogous pair of sites in the two cell lines. A site observed in only one cell line is shown with that cell line designated underneath. The ISRE (-) and the first exon (U) are shown. The subcloned fragment used as a probe (restriction sites and =) is indicated.

Article Snippet: WI38VA (ATCC CCL 75.1) or Daudi

Techniques: Comparison, Isolation, Marker, Electrophoresis

Journal: Phytomedicine

Article Title: GMI, a protein from Ganoderma microsporum , induces ACE2 degradation to alleviate infection of SARS-CoV-2 Spike-pseudotyped virus

doi: 10.1016/j.phymed.2022.154215

Figure Lengend Snippet: GMI downregulated expression of ACE2 in WI38–2RA cells in vitro and lung tissue of mice in vivo . (A) The viability of WI38–2RA cells treated with GMI was evaluated. Each GMI-treated group was normalized by the control group. (B) The ACE2 levels of WI38–2RA cells treated with GMI were evaluated by Western blot. (C-F) WI38–2RA cells were pretreated with DMSO/ddH 2 O (vehicle control) or MG132 (10 μM; C)/Bafilomycin A1 (BafA1; 20 nM; D)/dynasore (200 μM; E)/MβCD (20 mM; F) for 30 min, followed by incubation with GMI (0.6 μM) for 24 h and their ACE2 expression levels were evaluated. (G) Scheme for mouse receiving GMI via inhalation method. (H) The expressions of ACE2 protein in lung tissue were determined by Western blot. Tubulin was used as internal control. Each number (#1, #2, #3) represents the data collected from one single mouse. The data were representatives of more than three separated experiments and were presented as mean ± SDs; the error bars indicated SD. Significant differences were noted (*** p < 0.001).

Article Snippet: The normal human lung fibroblast WI-38 VA-13 subline 2RA (WI38–2RA) cells were purchased from the Bioresource Collection and Research Center of the Food Industry Research and Development Institute (BCRC, Hsinchu, Taiwan).

Techniques: Expressing, In Vitro, In Vivo, Control, Western Blot, Incubation

Journal:

Article Title: Selective killing of transformed cells by cyclin/cyclin-dependent kinase 2 antagonists

doi:

Figure Lengend Snippet: Preferential killing of transformed cells by peptidic cyclin/cdk2 antagonists. (A) The indicated cell lines were treated with the Tat-LDL peptide. Cell viability was measured by using an MTS assay. (B) WI38 and WI38/VA13 cells were treated with the indicated peptides, and cell viability was measured by MTS assay. (C). Phase-contrast (Left) and fluorescence microscopy (Right) of WI38/VA13 (Upper) and WI38 cells (Lower) treated with fluorescein-labeled Tat-LDL peptide.

Article Snippet: U2OS osteosarcoma cells, MDA-MB-435 breast carcinoma cells, HCT116 and SW480 colon carcinoma cells, WI38 human diploid fibroblasts, and the WI38 T-antigen-transformed subline WI38/VA13 were obtained from the American Type Culture Collection.

Techniques: Transformation Assay, MTS Assay, Fluorescence, Microscopy, Labeling