Figure 1 | Scientific Reports

Figure 1

From: A missense MT-ND5 mutation in differentiated Parkinson Disease cytoplasmic hybrid induces ROS-dependent DNA Damage Response amplified by DROSHA

Figure 1

SH-SY5Y cells and cybrids were differentiated to dopaminergic neuron-like cells. (A) SH-SY5Y cells were grown in differentiation media as described in Materials and Methods. Upper panels show the alternative splicing profile of PTBP2 exon 10 before (day 0) and after differentiation (day 12) (left) and the level of PTB and nPTB proteins (right). In the left panel, the upper band shows the full-length PTBP2 that includes exon 10 while the lower band shows the PTBP2 transcript produced by exon 10 skipping. At day 1, cells were transfected with control siRNA (Ctr) or with siRNA directed against PTBP1 mRNA. Lower panels show Western blot analyses of three neuronal differentiation markers, microtubule-associated protein 2 (MAP2), beta-3-tubulin (βIII tub) and tyrosine hydroxylase (TH), on whole SH-SY5Y cell lysates before (day 0) and after (day 12) differentiation. GAPDH was used as loading control. (B) PD61, PD63, PD67 cybrids and SH-SY5Y cells were grown in differentiation media. At day 12 total RNA was purified and the alternative splicing profile of PTBP2 exon 10 was analysed by RT-PCR (upper panel). Lower panel shows the Western blot analyses of PTB, nPTB and phosphorylated histone H2AX (γH2AX) before and after differentiation on the indicated cell lines. GAPDH, loading control.

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