Effects of double knockdowns of ASCC3 and genes involved in translation quality control on PF8503 toxicity.
Nadège Liaud
Max A. Horlbeck
Luke A. Gilbert
Ketrin Gjoni
Jonathan S. Weissman
Jamie H. D. Cate
10.1371/journal.pgen.1008057.g005
https://plos.figshare.com/articles/figure/Effects_of_double_knockdowns_of_ASCC3_and_genes_involved_in_translation_quality_control_on_PF8503_toxicity_/7853105
<p>(A) Lentiviral construct used to generate double knockdown cell lines. For each construct, the sgRNA targeting ASCC3 was placed after the human U6 (hU6) promoter, and a second sgRNA (scrambled or targeting ASCC2, NEMF, or HBS1L) was placed after the murine U6 (mU6) promoter. (B) PF8503 phenotype (Rho) obtained in competitive growth assays performed using double knockdown cell lines (grey), compared with the rho phenotype expected from the sum of phenotypes of individual knockdowns. Due to less efficient knockdown of ASCC3 in the dual-sgRNA context, the individual phenotype of ASCC3 knockdown was taken from the cell line expressing mU6-scrambled sgRNA-hU6-ASCC3 sgRNA pair. All other phenotypes were taken from the individual knockdowns in <b><a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1008057#pgen.1008057.g002" target="_blank">Fig 2A</a></b>. Experiments were carried out in biological triplicate for ASCC3-NEMF cell line and in 6 replicates for other cell lines, with mean and standard deviation shown.</p>
2019-03-15 17:39:07
compound
ubiquitin binding protein ASCC 2
translation quality control proteins Pelota
PCSK 9 translational inhibitor
LDL
genome-wide CRISPRi screen
HBS 1L
PF 8503 treatment
ribosome quality control pathways
ASCC 3 acts
PELO
stall protein synthesis