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. 2018 Sep 24;9(10):971. doi: 10.1038/s41419-018-1039-2

Fig. 2. Oncogenic Dbl-driven endocytosis derailment is modulated by GRP75.

Fig. 2

a Dbl-KD (D3) and NC SKOV-3 stable cell lines were transfected either with the GRP75-targeting shRNA Lentivirus (GRP75-KD), or with GRP75-pEGFP plasmids, or further treated with its chemical inhibitor MKT077 (40 µM, 12 h). After culture for 36 h, cells had Tfn-AF647, CTxB-AF647, Dextran-Rhodamine, scFv-αHS-AF647, and Tat/pGL3-YOYO-1 complexes added for 37 °C uptake as described in “Materials and methods”. The uptakes of each drug were determined by confocal imaging analysis, and representative images from three independent experiments are shown. Scale bar: 20 µm. The uptake variability of indicated transfections/treatments in single cell populations is shown in Supplementary Fig. 3 A–D; b GRP75 knockdown Cos-7 stable cell lines (GRP75-KD; G1, G2, G3) were produced by the CRISPR–CAS9 system. GRP75-KD (G3), NC Cos-7 stable cell lines, and MKT077-treated Cos-7 cells were transfected with either GST-proto-Dbl or GST-onco-Dbl plasmids. The incubation of fluorescent-labeled drugs and confocal-based uptake analysis were as described as above. The uptake variability in single cell populations is shown in Supplementary Fig. 3e–i