bacmid transfection agent promega fugene® hd  (Promega)

Journal: Human Genomics

Article Title: Zebrafish as a model to investigate a biallelic gain-of-function variant in MSGN1, associated with a novel skeletal dysplasia syndrome

doi: 10.1186/s40246-024-00593-w

Figure Lengend Snippet: In vitro transfection of HEK 293T cells with CMV:MSGN1 (WT)-FLAG-tag and CMV:MSGN1 p.(Arg125Leu)-FLAG-tag plasmids at different concentrations. A Immunofluorescence showing expression of tagged MSGN1 proteins (WT) or p.(Arg125Leu) after 48h of transfection. Corresponding experimental controls are shown in Additional file : Fig. S3A. B Quantification of MSGN1 localization in transfected cells. Fluorescence signals in a by single z-plane were visualized by confocal laser-scanning microscopy and subsequently signal intensity was measured in single cells (cyto: cytoplasm; nuc: nucleus) and outside of cells (bg: background). Graphs show signal intensity measurements of the RFP channel (MSGN1 Flag-Tag) of 30 cells per experimental group and 30 background positions. Signal intensity measurements of the corresponding DAPI channel for nucleus identification are given in Additional file : Fig S3B. Values are given in Additional file : Excel file S1

Article Snippet: 0.5 µg and 1.0 µg plasmid DNA per 100 µl total transfection volume was used with 2 to 3 µl FuGene HD transfection agent per well (Promega, Madison, USA, product-nr. E2311).

Techniques: In Vitro, Transfection, FLAG-tag, Immunofluorescence, Expressing, Fluorescence, Confocal Laser Scanning Microscopy