ENCBS458AAA / cell line

Status: released
Term name: K562
Term ID: EFO:0002067
Summary: Homo sapiens K562 cell line stably expressing C-terminal eGFP-tagged ATF3
Description: K562 cell line stably expressing a C-terminal LAP-tag containing eGFP, fused to ATF3-L
Culture start date: 2014-10-29
Culture harvest date: 2014-11-21
Passage number: 6

Accession	Category	Purpose	Method	Nucleic acid delivery method	Site
ENCGM032GSF	insertion	tagging		stable transfection	Target ATF3-human Coordinates hg19 chr1:212658547-212847095

Caption excerpt: Western blot analysis of nuclear lysates prepared from a K562 stable cell line…

Caption: Western blot analysis of nuclear lysates prepared from a K562 stable cell line expressin eGFP-ATF3-L using the Roche anti-GFP 11814450001 antibody. The expected MW is 54kDa.
Submitted by: Mark Murphy
Lab: Kevin White, UChicago
Grant: U54HG006996

Caption: immunoblot of GFP tagged ATF3-L, expressed in K562 cells

Caption excerpt: eGFP-ATF3 tagged cells were stained with a nuclear dye and sorted by FACS.…

Caption: eGFP-ATF3 tagged cells were stained with a nuclear dye and sorted by FACS. Intensities in the GFP channel are on the x-axis, nuclear stain on the y-axis. The boxed regions define a range of intensities in both channels corresponding to GFP+ and GFP++ for the high GFP and highest GFP expression cell populations, respectively. 2389 cells were counted in total, with 2.34% in the GFP+ region and 0.08% in the GFP++ regions respectively. The cells in these regions were further counted for GFP and nuclear signal colocalization and reported as a percentage of GFP+ and GFP++ cells localized to the nucleus: 56.9%. This enriched stable cell line population is then isolated, expanded and stored as frozen aliquots for downstream experiments.
Submitted by: Jay Rehm
Lab: Kevin White, UChicago
Grant: U54HG006996

Caption: eGFP-ATF3 tagged cells were stained with a nuclear dye and sorted by FACS.

Caption: eGFP-ATF3 tagged cells were stained with a nuclear dye and sorted by FACS.

Caption excerpt: Western blot analysis of nuclear lysates prepared from a K562 stable cell line…

Caption: Western blot analysis of nuclear lysates prepared from a K562 stable cell line expressing eGFP-ATF3 using the Roche anti-GFP 11814450001 antibody. The expected MW is 54kDa.
Submitted by: Jay Rehm
Lab: Kevin White, UChicago
Grant: U54HG006996

Caption excerpt: eGFP-ATF3-L tagged cells were stained with a nuclear dye, Vybrant DyeCycle…

Caption: eGFP-ATF3-L tagged cells were stained with a nuclear dye, Vybrant DyeCycle Ruby Stain from Life technologies, and sorted by FACS. Shown are the cells from the GFP+ fraction quantified for their GFP and nuclear signal colocalization.
Submitted by: Mark Murphy
Lab: Kevin White, UChicago
Grant: U54HG006996

Caption excerpt: eGFP-ATF3-L tagged cells were stained with a nuclear dye, Vybrant DyeCycle…

Caption: eGFP-ATF3-L tagged cells were stained with a nuclear dye, Vybrant DyeCycle Ruby Stain from Life technologies, and sorted by FACS. Shown are the cells from the GFP++ fraction quantified for their GFP and nuclear signal colocalization.
Submitted by: Mark Murphy
Lab: Kevin White, UChicago
Grant: U54HG006996

Caption excerpt: eGFP-ATF3-L tagged cells were stained with a nuclear dye, Vybrant DyeCycle…

Caption: eGFP-ATF3-L tagged cells were stained with a nuclear dye, Vybrant DyeCycle Ruby Stain from Life technologies, and sorted by FACS. Shown are the cells from the GFP+ fraction quantified for their GFP and nuclear signal colocalization.
Submitted by: Mark Murphy
Lab: Kevin White, UChicago
Grant: U54HG006996

Description: Protocol for generating GFP tagged BAC transgenic lines in K562.