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. 2016 Nov 17;140(1):49–67. doi: 10.1093/brain/aww272

Figure 4.

Figure 4

Immunoblotting studies obtained for mutant γ2L subunits. Wild-type or mutant γ2L subunits were cotransfected with α1β2 subunits into HEK293T cells. (A) Total cell lysates were collected, analysed by SDS-PAGE and blotted by anti-γ2 and anti-ATPase antibodies. In this representative western blot, NS = non-specific control. (B) Band intensity of γ2L subunits was normalized to the ATPase signal (n = 4, mean ± SEM). Both the lower and higher molecular mass bands were included. (C) Surface protein samples were collected through biotinylation and probed by anti-γ2 and anti-ATPase antibodies. A representative western blot is presented. (D) Band intensities of γ2L subunits were normalized to the ATPase signal (n ≥ 4, mean ± SEM). One-way ANOVA followed by Dunnett’s multiple comparison test were used to determine significance. *P < 0.05, compared with wild-type condition. (E) Surface proteins of HEK293T cells co-expressing α1β2γ2L, α1β2γ2L(N105Q), α1β2γ2L(I107T) or α1β2γ2L(N105Q/I107T) subunits were collected and probed with anti-γ2 and anti-ATPase antibodies. A representative western blot is presented. WT = wild-type.