Supplementary Figure 6: Characterization of IgSF11-knockdown and IgSF11 rescue constructs.
From: Synaptic adhesion molecule IgSF11 regulates synaptic transmission and plasticity
(a) Acute knockdown of IgSF11 expression in heterologous cells. HEK293T cells were doubly transfected with rIgSF11 (rat IgSF11) and sh-vec (empty vector), sh-IgSF11#1, or sh-IgSF11#1* (a mutant incapable of knocking down IgSF11) for two days, and the cell lysates were characterized by immunoblotting for IgSF11 and α-tubulin (loading control). n = 3, * p < 0.05, ** p < 0.01, ns, not significant, one-way ANOVA. Full-length blots are presented in Supplementary Figure 17d.
(b) Acute knockdown of IgSF11 expression by an independent IgSF11 knockdown construct (sh-IgSF11#2). HEK293T cells were transfected with doubly transfected with IgSF11 and sh-vec (empty vector) or sh-IgSF11#2 for two days, and the cell lysates were characterized by immunoblotting for IgSF11 and α-tubulin (loading control). n = 3, *** p < 0.001, Student’s t-test. Full-length blots are presented in Supplementary Figure 17e.
(c and d) Characterization of an sh-IgSF11#1-resistant IgSF11 expression construct. HEK293T cells were doubly transfected with IgSF11 (rat) + sh-vec, IgSF11 + sh-IgSF11#1, or sh-IgSF11#1-resistant IgSF11 (WT or ΔC3) + sh-IgSF11#1, followed by immunoblotting with IgSF11 antibodies. Three independent experiments were performed. Full-length blots are presented in Supplementary Figure 17f,g.
