Of note, we failed to detect in the mutant cells expression of the putative shorter RIPK1 product (data not shown), which may originate from an internal promoter [7].
While expression of full-length mTor failed to complement yeast tor1 or tor2 mutant cells, expression of Tor1-mTor and Tor2-mTor hybrids supported Tor-dependent, rapamycin-sensitive growth of yeast cells [31].
In contrast, metabolic gene expression was strongly decreased in wildtype cells upon LPS treatment, while in Tyk2 mutant cells the expression of these genes remained relatively unchanged, which exaggerated differences already present at the basal level.
Strains EEY6-2 and EEY9 correspond to mutant cells lacking expression of Vps23p and Snf7p (also known as Vps32p), respectively.
We show that UME6 was down-regulated in efg1Δ mutant cells, however, expression of UME6 was restored by forced overexpression of EED1.
In fact, in our study, mutant cells lacking expression of GRASP, which is required for unconventional secretion of an acyl coenzyme A binding protein in S. cerevisiae [39] and Dictyostelium discoideum [4], showed a decreased content of extracellular vesicles, in comparison with WT cells.
We have now carefully documented the mitotic progression in His C mutant cells upon expression of string and we found only minor mitotic defects.
We found that RANKL induced the expression of IFNβ in both WT and Optn D477N/D477N mutant cells, but the expression levels were significantly lower in mutant cells compared to those observed in WT.
Follicle cell clones mutant for awd show strong expression of FasIII after stage 6, indicating that they are defective in terminal differentiation (awd mutant cells lacking GFP expression in Figure 2C).
Most notably, mutant cells acquire gene expression that is characteristic of adjacent somatic cells, indicating that Prdm14 is crucial for PGC specification by promoting expression of germ cell genes while repressing somatic genes.
Therefore, loss-of-function screens with mutant cells that lack expression of a particular gene are difficult in diploid cells, because in most cases both alleles of a gene must be knocked out to result in a phenotype.
