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. 2018 Jul 16;2(3):NS20180141. doi: 10.1042/NS20180141

Figure 4. TAF1-depletion and transfection with TAF1 variant, p.Ser1600Gly significantly alters significantly neuronal ion expression at both the gene and protein levels.

Figure 4

(A) SH-SY5Y cells were transfected with a commercially available TAF1-specific siRNA duplex or a scramble non-targetting control duplex. β-ACTIN (HKG), TAF1, and CACNA1G gene mRNA expression were analyzed using qRT-PCR. All gene expression were calculated relative to B-ACTIN. (B) Protein lysates were subjected to Western blot analysis with antibodies directed against TAF1, and CACNA1G; β-Actin served as a loading control. (C) SH-SY5Y cells were transfected with a TAF1 wild-type plasmid expression vector or a TAF1 variant p.Ser1600Gly plasmid expression vector. β-ACTIN (HKG), TAF1, and CACNA1G gene mRNA expression was analyzed using qRT-PCR. TAF1 and CACNA1G gene expression of the TAF1 variant p.Ser1600Gly was calculated respective to the expression of the TAF1 wild-type plasmid samples. (D) SH-SY5Y cells were transfected with a TAF1 wild-type or TAF1 p.Ser1600Gly expression vector, protein isolated and subjected to Western blot analysis with antibodies directed against TAF1, CACNA1G, or β-Actin (loading control). For protein analysis, band intensities were quantitated from the 16-bit digital image by densitometry in ImageJ (NCBI) and are shown normalized to β-Actin control. All data are representative of three independent experiments with each sample repeated in triplicate.