Extended Data Figure 5: Impairment of mitochondrial translation due to loss of SHMT2.
From: Mitochondrial translation requires folate-dependent tRNA methylation
a, SDS–PAGE of [35S]methionine-labelled mitochondrially translated proteins in wild-type (lane 1) and two SHMT2-knockout (lane 2 and 3) HEK293T cell lines. Decreased synthesis of COX1 and COX2/3 are evident upon short exposure and reduced synthesis of ND5 and ND6 is more easily visualized upon longer exposure. b, Absorbance at 254 nm upon sucrose gradient fractionation of cell lysates digested by micrococcal nuclease (Fig. 3a). Fractions corresponding to 4 and 5 were collected for mitochondrial ribosome enrichment as shown on the matched immunoblot for mitochondrial ribosome subunit MRPL11. c, Read length distribution (top) and read length-dependent sub-codon read phasing (bottom) across the 13 mitochondrial protein-coding transcripts. Data in c are based on the mitochondrial ribosome profiling experiment in Fig. 3, and represent the mean of two technical replicates of two independent samples.
