Fig. 5

Mtf1 regulates HD-related processes

a Left: Immunofluorescence for MTF1 in Q128_Mtf1 cells, representative of n = 3 experiments. Nuclei were stained with DAPI. Scale bar, 30 μm. Right: Violin plot showing the fraction of MTF1 nuclear signal over the total signal from 1 experiment. b Heatmap showing the effect of Mtf1 overexpression on DEGs between Q128 and Q15 cells. MTF1 rescued 138 genes (p-value ≥ 0.05 between Q128_Mtf1 and Q15 and log₂ FC > |0.5| between Q128_Mtf1 and Q128). Mean expression of n = 6 (Q15), n = 7 (Q128), and n = 6 biological replicates (Q128_Mtf1). Z-score calculated on row-scaled expression values. c Processes altered by mHTT and enriched among genes rescued, or not rescued, by MTF1. P-values were calculated by two-tailed Fisher Exact test using Enrichr software. d RNAseq on Q128_Mtf1 and Q128_mCherry cells. DOWN-regulated (log₂ FC  <  −0.5 and p-value  <0.05) and UP-regulated (log₂ FC  > 0.5 and p-value  <0.05, dashed lines) genes are indicated in blue and red, respectively. P-values were calculated with two-tailed Wald Test with no adjustment. N = 7 (Q128_mCherry) and n = 6 biological replicates (Q128_Mtf1 cells). e Measurement of cell death by PI uptake and Flow Cytometry. Bars indicate the mean of 2 independent experiments, shown as dots. Representative profiles are shown on the right. f Measurement of ROS production by H₂DCFDA staining. Bars indicate the mean of 2 independent experiments shown as dots. Representative profiles are on the right. g Cellular uptake of metals in Q15 and Q128 cell lines. Left: heatmap of mean-normalised intracellular concentrations. Measurements of total intracellular (middle) and extracellular (right) amount of Cadmium in Q15_HTT and Q128_HTT cells. Fresh medium served as negative control. Mean and SEM of at least 3 biological replicates, shown as dots. P-values calculated with the unpaired two-tailed Mann-Whitney U test. h Top: consensus sequence recognised by MTF1 from Jaspar database, identified in MREs. Bottom: gene tracks for Mt1 and Mt2, showing the expression levels in Q128_Mtf1 and Q128_mCherry cells; one representative biological replicate of RNA sequencing data is shown for each cell line.