Figure 6.

Thoc1 deficiency compromises protein synthesis and cell growth. A) The effect of pThoc1 depletion on the rate of protein translation was assayed in viable AdCre (cre)- or AdGFP (gfp)-infected PrT of the indicated genotypes by ³⁵S-methionine incorporation. The data show the ratio of ³⁵S incorporated into trichloroacetic acid-precipitated protein vs total cellular ³⁵S during a pulse label. Each data point is from an independent experiment using two different PrT isolates for each genotype. The bars represent the mean and standard deviation. The asterisk indicates a statistically significant difference (t test P < .001). B) Autophagy in PrT was characterized by western blot analysis of LC3B. PrT of the indicated genotypes were infected with AdCre (cre), AdGFP (gfp), or treated with chloroquine (cq) for one hour prior to extraction. Increased conversion of the LC3B-I to LC3B-II is apparent in pThoc1-depleted PrT. The blot is representative of multiple experiments with independent PrT isolates. Actin is the protein loading control. C) PrT of the indicated genotypes were treated with AdCre (cre) or AdGFP (gfp) and the diameter of viable cells measured using a Vi-cell Coulter counter. The histogram shows cumulative data for three to four independent experiments using two to three different PrT isolates for each genotype. All statistical tests were two-sided.