Journal: Neural Regeneration Research

Article Title: Overexpression of cytoglobin gene inhibits hypoxic injury to SH-SY5Y neuroblastoma cells

doi: 10.3969/j.issn.1673-5374.2013.23.010

Figure Lengend Snippet: Transfected SH-SY5Y-GFP-cytoglobin cells (fluorescence microscopy, × 100). SH-SY5Y cells were transfected with pAcGFP1-C1-cytoglobin using Lipofectamine 2000. The successfully-transfected cells showed green fluorescence. GFP: Green fluorescent protein.

Article Snippet: SH-SY5Y cells were obtained from the Cancer Institute of the Chinese Academy of Medical Sciences. pAcGFP1-C1-cytoglobin was obtained from Takara Biotechnology (Dalian) Co., Ltd., Liaoning Province, China.

Techniques: Transfection, Fluorescence, Microscopy

Journal: Nature Communications

Article Title: Directed natural evolution generates a next-generation oncolytic virus with a high potency and safety profile

doi: 10.1038/s41467-023-39156-3

Figure Lengend Snippet: a A modified plaque formation assay was performed and cells were imaged with a fluorescence microscope 48 hours after infection (MOI = 0.5). Scale bars, 500 μm. The scale bars in the magnified images represent 100 μm. b Plaques were counted with a fluorescence microscope 48 hours after infection. Data points represent mean plaque number per well ± SD, for n = 3 biological replicates. P < 0.0001 was determined by Two-way ANOVA relative to M1-GFP. c HCT-116 cells were incubated with M1-GFP and M1-E2M at 4 °C for 1 hours. Viral RNA was quantified by qRT-PCR and presented as mean ± SD, for n = 3 biological replicates. P < 0.0001 was calculated with Two-tailed unpaired t-test. d Western blot analysis of M1-GFP and M1-E2M incubated with MXRA8-His bound to His-Tag Mouse mAb Sepharose Beads. Precipitated viral particles were detected using an anti-E1 mAb (left). Quantification of E1 expression is shown (right). Graph bars represent mean densitometry of E1 normalized to the densitometry of MXRA8 ± SD, for n = 3 biological replicates. P = 0.0060 was calculated with Two-tailed unpaired t-test. e Time course of the binding between M1 viral particles and the MXRA8 protein, as determined via BLI. f , g HeLa, HeLa-Mxra8, Hs 578 T and Hs 578T-ΔMxra8 cells were treated with M1-GFP or M1-E2M. The infection rate was determined by flow cytometry 48 hours after infection. Graph bars represent mean infection rate % ± SD, for n = 3 biological replicates. Statistical significance was calculated using Two-way ANOVA with Sidak’s multiple comparisons test relative to M1-GFP. Adjusted P values are: f vector P = 0.6199; Mxra8 P = 0.0002; g vector, P = 0.0010; ΔMxra8 P = 0.2713. n.s.: no significance, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Source data are provided as a Source Data file.

Article Snippet: M1-GFP and M1-E2M (1×10 7 PFU) virus particles, 1 μg of His-Mxra8 (Sino Biological), and His-Tag Mouse mAb Sepharose Beads (Cell Signaling Technology) were incubated overnight at 4 °C in TBS containing 10 mM CaCl 2 .

Techniques: Modification, Plaque Formation Assay, Fluorescence, Microscopy, Infection, Incubation, Quantitative RT-PCR, Two Tailed Test, Western Blot, Expressing, Binding Assay, Flow Cytometry, Plasmid Preparation