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. 2021 Apr 24;296:100707. doi: 10.1016/j.jbc.2021.100707

Figure 3.

Figure 3

Direct interaction of CtIP with GDCR8.A, the direct interaction between CtIP and DGCR8 was shown using a GST pull-down assay. B, pull-down experiments were performed by incubating lysates prepared from insect cells infected with Flag-DGCR8 baculoviruses with different purified GST-CtIP fragments. C, pull-down experiments were performed by incubating lysates prepared from insect cells infected with Flag-CtIP baculoviruses with different purified GST-DGCR8 fragments. D, 293T cells were cotransfected with CtIP-VN and DGCR8-VC. Reconstituted YFP fluorophore was visualized via total internal reflection fluorescence microscope at 24 h after transfection. Top, schematic of BiFC system. Bottom, representative fluorescence images of CtIP and DGCR8 interaction in cells. The scale bars represent 100 μm. E, detection of CtIP–DGCR8 colocalization was performed using a PLA probe in U2OS cells. Representative PLA foci (green) are shown. The scale bars represent 10 μm. CtIP, C-terminal-binding protein–interacting protein; GDCR8, DiGeorge syndrome critical region gene 8; GST, glutathione-S-transferase; U2OS, human osteosarcoma cell line; PLA, proximity ligation assay; VC, Venus C-terminal fragment; VN, Venus N-terminal fragment.