Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2020 Jul 6;21(13):4781. doi: 10.3390/ijms21134781

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Immunofluorescence analysis of CFTR protein in triton permeabilized FRT-CFTR^W1282X cells and quantification of the immunofluorescence. FRT-CFTR^W1282X cells were transfected with the plasmids encoding the indicated gRNAs and dCAS13b/ADAR2_DD (300 ng total DNA). Untransfected FRT-CFTR^W1282X cells (NT) were used as negative control. FRT-CFTR^W1282X cells treated with G418 and FRT-CFTR^WT cells were used as a positive control. (A) The CFTR protein was revealed by the primary antibody mAb570, followed by a secondary antibody anti-mouse-FITC conjugated (green, Sigma). Nuclei (blue) were DAPI stained. Images were taken at 63x magnification on a ZEISS microscope equipped for epifluorescence. (B) The quantification is relative to the replicate representative of the results. The error bar represents the standard error of the mean (SEM). The quantification of CFTR signal was done manually by using Fiji software. The cell contour was drawn by using a cell membrane marker. The background was subtracted and the integrated signal intensity in the selected area (one cell) was measured.

Immunofluorescence analysis of CFTR protein in triton permeabilized FRT-CFTR^W1282X cells and quantification of the immunofluorescence. FRT-CFTR^W1282X cells were transfected with the plasmids encoding the indicated gRNAs and dCAS13b/ADAR2_DD (300 ng total DNA). Untransfected FRT-CFTR^W1282X cells (NT) were used as negative control. FRT-CFTR^W1282X cells treated with G418 and FRT-CFTR^WT cells were used as a positive control. (A) The CFTR protein was revealed by the primary antibody mAb570, followed by a secondary antibody anti-mouse-FITC conjugated (green, Sigma). Nuclei (blue) were DAPI stained. Images were taken at 63x magnification on a ZEISS microscope equipped for epifluorescence. (B) The quantification is relative to the replicate representative of the results. The error bar represents the standard error of the mean (SEM). The quantification of CFTR signal was done manually by using Fiji software. The cell contour was drawn by using a cell membrane marker. The background was subtracted and the integrated signal intensity in the selected area (one cell) was measured.