Skip to main content
. 2020 Mar 4;295(16):5229–5244. doi: 10.1074/jbc.RA120.012628

Figure 3.

Figure 3.

PKC-induced DAT internalization in females is limited to ventral striatum and requires Rit2. Conditional Rit2 silencing in DA neurons and ex vivo striatal slice biotinylation. Female Pitx3IRES2-tTA/+ mouse VTA were bilaterally injected with either AAV9-TRE-eGFP or -shRit2. Brains were harvested 4–5 weeks postinjection, and DAT surface expression was measured in ex vivo striatal slices by surface biotinylation as described under “Experimental procedures,” following treatment with or without 1 μm PMA for 30 min at 37 °C. Representative blots are shown in the top of each panel, and average data are presented at the bottom of each panel. DAT surface levels are expressed as percentage of total DAT ± S.E. (error bars), n = 5–8 slices from n = 3 independent mice/virus. A, dorsal striatum. Neither PKC activation nor Rit2-KD had an effect on DAT surface expression (two-way ANOVA: interaction: F(1, 22) = 0.051, p = 0.82; drug: F(1, 22) = 0.58, p = 0.46; virus: F(1, 22) = 0.18, p = 0.68). B, ventral striatum. PKC activation and Rit2-KD significantly decreased DAT surface expression, and Rit2-KD blocked PKC-stimulated DAT internalization (two-way ANOVA: interaction: F(1, 18) = 4.54, p = 0.047; drug: F(1, 18) = 4.25, p = 0.054; virus: F(1, 18) = 7.28, p = 0.015. Sidak's multiple comparisons test: eGFP(veh) versus eGFP(PMA): *, p = 0.04; eGFP(veh) versus shRit2(veh): *, p = 0.02; shRit2(veh) versus shRit2(PMA): p > 0.99).