Figure 7
HAP1 and TBP-FL, -ΔQ, or -Q50 co-localization assays in COS-7 and Neuro-2a cells. (A) Top two rows show cells transfected with a single expression plasmid: CMVp-GFP-TBP-FL (at left) and CMVp-DSRed2-HAP1-B155–582 (at right). GFP-TBP was exclusively nuclear and RFP-HAP1 assembled into cytoplasmic STLBs. Bottom two rows show co-transfected cells containing GFP-TBP and RFP-HAP1. Upon co-expression with HAP1, a portion of GFP-TBP became localized to the cytoplasmic, RFP-HAP1-STLBs. (B) Top two rows show cells transfected with a single expression plasmid: CMVp-DSRed2-TBP-ΔQ (at left) and CMVp-DSRed2-TBP-Q50 (at right). Both RFP-TBP-ΔQ and RFP-TBP-Q50 proteins localized exclusively to the nucleus when expressed alone. Cells co-transfected with GFP-HAP1-A7–598 and either RFP-TBP-ΔQ (middle two rows) or RFP-TBP-Q50 (bottom two rows) showed altered localization of TBP away from the nucleus. RFP-TBP-ΔQ, which lacks the polyQ repeat, and RFP-TBP-Q50, which contains an expanded polyQ repeat, each were partially localized in the GFP-HAP1-STLBs.