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. 2020 Sep 18;13:573409. doi: 10.3389/fnmol.2020.573409

Figure 1.

Figure 1

Human TSC1 variants and expression in medial ganglionic eminences (MGE) primary cultures. (A) Schema depicting the lentiviral DNA vector and resulting lentivirus to express human TSC1 variants and Cre recombinase. (B) TSC1 variants expressed in HEK293T cells show increased levels of expression over endogenous HAMARTIN protein and migration at the correct molecular weight. (C,C’) E13.5 MGE primary cultures that were grown for 7 days and co-labeled for GABA and DAPI. E13.5 MGE primary cultures that were either transfected (D,D’) or transduced (E,E’) with the vector or virus depicted in (A) and co-labeled for the Cre-dependent reporter, tdTomato, and GABA. (F) Quantification of the proportion of DAPI+ cells that express GABA in MGE cultures. (G) Quantification of the proportion of tdTomato+ cells that are GABA+. Yellow arrows denote non-co-labeled cells while white arrows denote co-labeled cells. Data are expressed as the mean ± SEM. Data were collected from three biological replicates (transplants) for all groups. Abbreviations: WB, western blot; KD, kilodalton. Scale bars in (C’,E’) = 100 μm. ****p < 0.0001.