Skip to main content
Fig. 4 | Applied Biological Chemistry

Fig. 4

From: Escherichia coli methionine-tRNAi/methionyl tRNA synthetase pairs induced protein initiation of interest (PII) expression

Fig. 4

Comparison of enhanced green fluorescent protein (EGFP) mRNA expression following the initiation of the translational reading frame induced using non-formyl methionine Escherichia coli initiator tRNA (Met-tRNAi) from E. coli methionyl tRNA synthetase (EcMRS) and human methionyl tRNA synthetase (HMRS). A EGFP mRNA levels were quantified using quantitative real-time polymerase chain reaction (qRT-PCR) and normalized to GAPDH in the presence of E. coli Met-tRNAi/EcMRS pairs (blue bar) in HeLa cells. B Effect of E. coli Met-tRNAi with EcMRS on HeLa cell viability. Cell survival was measured using cell assays with specific concentrations (1, 5, and 10 nM) of E. coli Met-tRNAi/EcMRS pairs (blue bars). The effect of non-formylated L-methionine (L-Met) on HeLa cell viability was determined via qRT-PCR analysis of EGFP mRNA expression. HeLa cells were treated with (red bar) or without L-Met (orange bar) for 24 h. Cell viability was determined using CCK8 assays and calculated relative to that of the control HeLa cells (red bar). Data are presented as the mean ± SEM of three independent experiments. (C) qRT-PCR (right) and dot plots showing EGFP expression (left). HeLa cells were treated with E. coli Met-tRNAi/HMRS (green bar) for 24 h. The qRT-PCR results were normalized to GAPDH. Bars represent the mean ± standard deviation of three independent experiments. Dot plots for EGFP expression in a L-Met-free medium (orange line), HeLa cells only (red line), E. coli Met-tRNAi/EcMRS pairs (blue line) and E. coli Met-tRNAi/HMRS pairs (green line) are shown

Back to article page