Figure 4
Compound treatment increased α-syn mRNA and protein levels in SH-SY5Y wildtype cells. (A) Expression fold change of SNCA mRNA measured with RT-qPCR. (B) Protein levels were determined by ICW and (C) WB in SH-SY5Y wild type cells. RT-qPCR data were normalized to three housekeeping genes and DMSO control, in triplicates, respectively. The Boxplot diagrams for (B) ICW represent the normalized mean of four 96-well plates with six to seven repetitions per compound, respectively. (C) For WBs six (VPA n = four) repetitions were conducted. Boxplot diagrams represent 5–95 percentile. (C, lower) Representative WB. Compounds were applied at a final concentration of 25 µM (Clo 12.5 µM) for 24 h. VPA was used as a positive control at a concentration of 600 µM. *p < 0.1 **p < 0.01, ***p < 0.001, ****p = 0.0001. Full-size WB of DMSO and Clo see Supplementary Fig. S4A, DMSO and Coni see Supplementary Fig. S4B, DMSO and Emo see Supplementary Fig. S4C and DMSO and VPA see Supplementary Fig. S4D. ICWs (B) and WB (C) membranes were imaged with the LI-COR Odysseys Clx (Model 9140; S/N CLX-0554) and signals were quantified using the Image Studio software 4.0.21.
