Skip to main content
. 2020 Mar 17;48(17):9762–9786. doi: 10.1093/nar/gkaa148

Figure 7.

Figure 7.

The knockdown of uS11m phenocopies the loss of YBEY. (A) High-resolution 10–40% glycerol gradient analysis of YBEY+ and YBEY KO total cell lysates followed by northern blotting shows a decreased association of the MT-ND3 mRNA and mt-tRNAMet with the mitochondrial SSU of YBEY KO cells. (B) Quantitative analysis of the association of extramitoribosomal MT-ND3 mRNA with the SSU in the YBEY+ and YBEY KO cells. Means ± SEM for n = 3 independent cell lines are shown; P-value, two-tailed Welch's test. (C) Metabolic [35S]-methionine labelling of mitochondrial translation products reveals a strong decrease of protein synthesis upon transient uS11m knockdown. A non-targeting control siRNA (‘Scrambled’) and two different uS11m-directed siRNA duplexes were used for HEK293T-REx cell transfection. The corresponding western blots confirm the uS11m knockdown and the concomitant depletion of mS37. VDAC1 is used as loading control. (D) High-resolution 10–40% glycerol gradient analysis of the total cell lysates from HEK293T-REx cells transiently transfected with control or uS11m-directed siRNAs followed by northern blotting shows a decreased association of the MT-ND3 mRNA and mt-tRNAMet with the mitochondrial SSU upon uS11m knockdown.