Journal: bioRxiv

Article Title: Unleashing a Novel Function of Endonuclease G in Mitochondrial Genome Instability

doi: 10.1101/2021.05.27.445952

Figure Lengend Snippet: A. Representative images of HeLa and HEK293T cells showing localization of BG4, the G4 binding antibody to mitochondria following immunofluorescence study. Nucleus is stained with DAPI (blue color), mitochondria with MitoTracker DR (red) and BG4 with Alexa-Fluor 488 (green). A merged image is shown with merge of red and green as depicted by Coste’s mask (colocalization is represented as a white dot). B, C. Quantitation showing colocalization of BG4 with MitoTracker indicated as dot plots. The colocalization was quantified using Mander’s colocalization coefficient (ImageJ software) analyzing a minimum of 100 cells as red over green (B) and green over red (C). D. Representative image of rho(0) cell showing localization of BG4 to mitochondria following immunofluorescence as investigated in panel A. E. Quantitation showing comparison of colocalization of BG4 between HeLa cells and rho(0) cells is shown as dot plots. The colocalization was quantified using Mander’s colocalization coefficient analyzing a minimum of 50 cells as red over green. F. BG4 bound mtDNA was purified after reverse crosslinking and used for real-time PCR using primers derived from different regions of the mitochondrial genome, which include 5 G-quadruplex forming regions and 10 random regions. Input DNA served as template control. No antibody control was also used. Bars in blue (first 5) are for G-quadruplex forming regions, while in green (last 10) are for random regions. Y-axis depicts threshold Ct value obtained following real time PCR for each primer. Error bar represents mean ± SEM. G. Agarose gel profile showing the amplification of Input DNA (left panel) and BG4 pull down DNA (right panel). “M” denotes 100 bp ladder.

Article Snippet: HeLa (human cervical cancer), MEF (mouse embryonic fibroblast) and HEK293T (human embryonic kidney epithelial cell line) were purchased from National Centre for Cell Science, Pune, India.

Techniques: Binding Assay, Immunofluorescence, Staining, Quantitation Assay, Software, Comparison, Purification, Real-time Polymerase Chain Reaction, Derivative Assay, Control, Agarose Gel Electrophoresis, Amplification

Journal: Oncology Letters

Article Title: Identification of a peptide specifically targeting ovarian cancer by the screening of a phage display peptide library

doi: 10.3892/ol.2016.4549

Figure Lengend Snippet: Immunofluorescence assay of phage clone P2 binding HO-8910, HeLa and CHO cells. The selected phage clone P2 was added individually to the cells and then incubated with anti-M13 antibody, FITC-labeled antibody and PI. The results were visualized using a fluorescence microscope. The results show that P2 could bind to HO-8910 cells but not to HeLa and CHO cells (control cells). Magnification, ×200. PI, propidium iodide; FITC, fluorescein isothiocyanate.

Article Snippet: The HO-8910 human ovarian cancer cell line, and HeLa cervical cancer cell line, were purchased from Jilin Baili Biotechnology Co., Ltd. (Changchun, China).

Techniques: Immunofluorescence, Binding Assay, Incubation, Labeling, Fluorescence, Microscopy, Control