Figure 5.

m¹A safeguards mRNAs during heat shock. (A) Schematic depiction of the m¹A motif-containing reporter. UTR, untranslated region. The position of the motif on the transcript is indicated. The nucleotide sequence of the motif is shown in Supplementary Figure S5B. (B) Accumulation of UbE from WT-UbE and MUT-UbE transcripts during 3 h-long inhibition of proteasomal degradation with 20 μM MG132 (+MG132). One representative anti-EGFP western blot from three independent experiments is shown. GAPDH was used as loading control. #, not significant difference; two-tailed t-test; N = 4 independent experiments (mean + SD). (C) Accumulation of UbE protein from WT-UbE and MUT-UbE transcripts during recovery after heat shock in the presence of actinomycin D to block synthesis of new mRNA molecules. UbE amount at time point 0 was subtracted from all values during recovery. One representative anti-EGFP western blot is shown. GAPDH was used as loading control. ***P < 0.001, **P < 0.01, two-tailed t-test; N = 4 independent experiments (mean + SD).