Fig. 2: Acetylome profiling during neuronal differentiation of controls and RTS patients hiPSCs. | Communications Biology

Fig. 2: Acetylome profiling during neuronal differentiation of controls and RTS patients hiPSCs.

From: Transcriptome and acetylome profiling identify crucial steps of neuronal differentiation in Rubinstein-Taybi syndrome

Fig. 2

a Schematic overview of the samples used and steps of analysis. hiPSCs were derived from primary fibroblasts from two healthy donors (controls) and four patients affected with RTS carrying the same CREBBP mutation: c.3832 C > T/p.Glu1278Lys. All hiPSCs clones were differentiated into cortical and pyramidal neurons. Acetylome analysis by mass spectrometry (LC–MS/MS) focused on core histones proteins (H2A, H2B, H3, and H4) and gene expression analysis by high-throughput RNA-sequencing was performed at three time points during the differenciation process (D10, D20, and D60). For each condition, two biological replicates corresponding to two independent differentiation experiments were used. b, c Kinetics of histone acetylation for H2A, H2B, H3, and H4 during neuronal differentiation at D10, D20, and D60 in controls (b) and RTS patients (c). The log2 ratio for each acetylation site of the indicated histones is shown for each time point. An acetylation site is considered hypoacetylated with a log2ratio ≤ 1 and hyperacetylated with a log2ratio ≥ 1 and the position is labeled on each graph. D10 was chosen as the reference value for the analysis. d–g Mapping of differential analysis of acetylome of CBP-dependent histone marks during neuronal differentiation in RTS patients compared to controls. Heatmaps of the acetylome during neuronal differentiation. The heatmaps display the log2 ratios for acetylation sites on the protein core histones H2A (d), H2B (e), H3 (f), and H4 (g) for each RTS patient (RTS1–RTS4) compared to controls at the 3 time points of the neuronal differentiation (D10, D20 and D60). An acetylation site is considered hypoacetylated with a log2ratio ≤ 1 and is displayed as a magenta gradient. Values with a log2ratio between −1 and 1 are shown in white. We considered a candidate histone mark altered in RTS as mark that is hypoacetylated in at least 3 out of the 4 RTS patients analyzed and for at least 1 out of the 3 time points.

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