In vitro GST pull-down The plasmids (Myc-PTEN or Myc-DUSP7) were transiently transfected into 293T cells. After transfection for 24 h, the cells were harvested and lysed in cell lysis buffer for Western and IP (Beyotime, P0013) containing protease inhibitor on ice. GST or GST-SPOPMATH...
GST pull-down assay demonstrating in vitro binding between MT-1X and FHL3.doi:10.1371/journal.pone.0093723.g003C. XinW. JinfengH. XinFu WenliangX. WenrongZou MinjiWang YuanyuanWang JiaxiXu Donggang
An in vitro GST pull-down assay showed that purified GST-Twa1 directly interacted with His-β-catenin (Supplementary information, Figure S7A). Immunoprecipitation analysis revealed that either endogenous or ectopically expressed Twa1 interacted with endogenous β-catenin (Supplementary information, Figure...
The in vitro radiosensitization effect of CDKL1 was evaluated using a neutral comet assay, γH2AX foci formation analysis, and a clonogenic cell survival assay. The protein‒protein interactions were confirmed via coimmunoprecipitation and GST pulldown assays. The regulation of PD-L1 by CDKL1 was...
GST Pull-Down Assay VASP cDNA was inserted into expression vector pGEX 6p-1, GST, or GST-VASP fusion proteins were purified fromEscherichia coli(BL21 DE3) using glutathione-Sepharose beads. GST pull-down assay was performed as previously described.19 ...
GST and GST-hnRNPK fusion protein extracts were absorbed on 100 μl (50% slurry) glutathione-Sepharose 4B beads (GE Healthcare), washed in 10 bed volumes of extraction buffer with 0.5% Triton X-100 and used as bait in a pulldown assay. Prior to in vitro binding the XRN2-His solution...
spot syndrome virus (WSSV)-VP37 and gill membrane proteins (GMPs) of Pacific white shrimp (Litopenaeus vannamei), the VP37 protein was expressed and purified, and a distinct 53 kDa VP37-binding protein band was identified in GMPs by virus overla...
We performed an in vitro pull-down assay to validate the interaction between OsUBC12 and OsSnRK1.1. OsSnRK1.1-His, OsUBC12-GST and GST alone were all detected in whole-cell lysates (Input). OsSnRK1.1-His was pulled down by OsUBC12-GST, but not by the GST-only control, suggesting th...
(b), in vitro pull-down assay (c), and BiFC (d). In Co-IP and BiFC assays, an irrelevant NAC protein (SNAC1) and the 1–80 amino acid (AA) truncated form of OsREM1.5 (OsREM1.51–80) were used as negative controls. In the pull-down assay, an irrelevant protein (OsPYL1) ...
GST pulldown assay using in vitro translated Myc-tagged Smad7 proteins was performed. Samples from the GST pulldown were resolved by SDS-PAGE and transferred to nitrocellulose membranes. Smad7 was detected by using αMyc antibody. Retroviral Infection—The pBABE, pBABE-SIRT1, pSUPERretro, and ...