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Increased Susceptibility to Cortical Spreading Depression in the Mouse Model of Familial Hemiplegic Migraine Type 2

Figure 3

In vitro expression and localization of α2-ATPase.

A. Immunoblot of HeLa cells transfected with pA2-wt-Myc (wt) and pA2-R887-Myc (R887) probed with anti α2-ATPase antibody. B. Calnexin (red) and α2 ATPase (probed with anti-myc; green) immunofluorescence staining of wt or R887 transfected Hela cells. R887 α2 ATPase mostly colocalises with the endoplasmic reticulum. In the right graphs, the scatter plot of red (ch1) vs. green (ch2) colocalization intensities. (wt parameters: Rr 0.369, R 0.6, Ch1:Ch2 0.999; R887 parameters: Rr 0.558, R 0.755, Ch1:Ch2 0.997). C. HeLa cells expressing wt or R887 variant of α2 ATPase are treated with the reversible proteasome inhibitor MG132 (10 µM for 4, 6 and 8 hrs). Blots are probed with anti- α2 isoform antibody, anti-ubiquitin antibody and anti-tubulin as loading control.

Figure 3