Fig. 3: Methylglyoxal suppresses Notch1 mRNA translation.

a–c E13.5 cortices were co-electroporated with nuclear EGFP and control or Glo1 (shGlo1) shRNAs and analyzed two days later. Images (a) and quantification (b) of EGFP + cells (green) also positive for Notch1 (red). White boxes in a show higher magnification at the bottom. Arrows denote double-labelled cells. n = 3 embryos each. c qRT-PCR analysis for Notch1 mRNA in FACS sorted EGFP+ cells from cortices as in a. n = 3 embryos each. d–i hNPCs were treated with DMSO or BBGD for 48 h. Images (d) and quantification (e) from western blots of hNPCs probed for NOTCH1 and ACTb. n = 3 experiments. f qRT-PCR analysis for NOTCH1 and NESTIN (NES) mRNAs in hNPCs. n = 3 experiments. g Polysome profile of hNPCs. Fractions corresponding to 40 S and 60 S ribosome subunits, the 80 S monosomes and polysomes are labelled. h Relative distribution of the NOTCH1 mRNA (left) and GAPDH mRNA (right) across all fractions measured by qRT-PCR. Each point corresponds to the value of each fraction normalized to the total NOTCH1 or GAPDH mRNA. i Quantification of the relative enrichment of NOTCH1 and GAPDH mRNAs in the heavy polysome fractions (#8–12). n = 3 experiments. j–l Dual-luciferase reporters containing the full-length (FL) or truncated Notch1 3ʹUTR (dARE) were co-electroporated into E13.5 cortices with control or Glo1 shRNAs. j Luciferase activity of the reporter. n = 6 embryos each. k Luciferase mRNA levels quantified by qRT-PCR. n = 3 embryos each. l Luciferase activity from the 3ʹUTR-dARE reporter. n = 8 embryos each. m, n hNPCs were transfected with dual-luciferase reporters and treated with DMSO or BBGD for 48 h. m Luciferase activity of the reporters.; n = 3 experiments. n Luciferase mRNA levels quantified by qRT-PCR. n = 3 experiments. Scale bars, 25 µm. Data are presented as mean values ± SEM and analyzed using two-tailed, unpaired students t-test. **p < 0.01, *p < 0.05, ns = p > 0.05. Source data and p-values are provided as a “Source Data file”.