RGS8 Antibody

Western Blot: RGS8 Antibody [NBP2-20153] -

Western Blot: RGS8 Antibody [NBP2-20153] - Overexpression of mutant ATXN2 in human SH-SY5Y cells recapitulates down-regulation of in vivo steady-state levels of Rgs8 in BAC-Q72 mice.Cells were transfected with plasmids encoding Flag-tagged cDNAs of human ATXN2 containing Q22 or Q58 or Q108 repeats. Forty-eight hrs post-transfection, cells were selected with hygromycin (40 µg/ml) for 5–7 days & hygromycin resistant cells were harvested as two aliquots. (A) Protein extracts were prepared from one aliquot & subjected to Western blot analyses to measure steady-state levels of RGS8. The blots were re-probed for beta-Actin as an internal loading control. (B) Quantitative RT-PCR analyses of synthesized cDNAs from the other aliquot demonstrate moderate reduction of RGS8 mRNA in cells expressing Flag-ATXN2-Q108. The data are means ± SD, *p<0.05. (C) Mutant ATXN2 specifically induces decrease of RGS8 expression. MYC-tagged RGS8 cDNA including 5’ & 3’ UTRs was cloned under the transcriptional control of the CMV promoter & transfected into short-term hygromycin selected SH-SY5Y cell lines expressing Flag-tagged ATXN2-Q22, -Q58 or -Q108. Forty-eight hrs post-transfection, levels of exogenous RGS8 are significantly decreased in cells expressing ATXN2-Q58 or -Q108 compared with cells expressing wild-type ATXN2-Q22. To control for equal transfection, we monitored levels of GFP, which was expressed as an independent cassette in the plasmid. Blots were re-probed for beta-actin as an internal loading control. The blot represents one of three independent experiments. Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pgen.1005182), licensed under a CC-BY license. Not internally tested by Novus Biologicals.