Journal: The Journal of Biological Chemistry

Article Title: Rab8 and TNPO1 are ciliary transport adaptors for GTPase Arl13b by interacting with its RVEP motif containing ciliary targeting sequence

doi: 10.1016/j.jbc.2023.104604

Figure Lengend Snippet: Rab8, TNPO1, and 13bCTS form a ternary complex. All cell lysates were from HEK293T cells. A, Rab8-TN interacts with Arl13b and TNPO1 in an RVEP motif-dependent manner. Bead-immobilized GST-Rab8-TN and GST-Rab8-QL were incubated with cell lysates transiently expressing RP2-GFP (positive control), Arl13b-WT-GFP, or Arl13b-RVEP-4A, and the proteins retained were analyzed by immunoblotting. B, a schematic diagram illustrating various recombinant His-tagged Arl13b fragments used in the below pull-down assays. C, Rab8-TN interacts with the C-terminal half of Arl13b. Bead-immobilized GST-Rab8-TN and GST-Rab8-QL were incubated with purified His-tagged N (His-AA1-245-GFP) or C-terminal half of Arl13b (His-AA193-428-GFP), followed by immunoblotting similar to (A). D, TNPO1 interacts with the C-terminal half of Arl13b in a Rab8-TN-dependent manner. Bead-immobilized GST-TNPO1 was incubated with purified His-tagged Rab8-TN, Rab8-QL, N (His-AA1-245-GFP), or C-terminal half of Arl13b (His-AA193-428-GFP), followed by immunoblotting similar to (A). E, TNPO1 interacts with 13bCTS. Bead-immobilized GST-TNPO1 was incubated with purified His-13bCTS-GFP or His-MBP-GFP (negative control), followed by immunoblotting similar to (A). F, TNPO1 interacts with 13bCTS in a Rab8-TN-dependent manner. Bead-immobilized GST-TNPO1 was incubated with purified His-tagged Rab8-TN, Rab8-QL, 13bCTS, or control, followed by immunoblotting similar to (A). ∗ indicates the specific band. Molecular weight markers are labeled to the right of all immunoblots.

Article Snippet: Cell culture and transfection RPE1 (hTERT-RPE1) and HEK293T cells were from American Type Culture Collection.

Techniques: Incubation, Expressing, Positive Control, Western Blot, Recombinant, Purification, Negative Control, Control, Molecular Weight, Labeling

Journal: Cell reports

Article Title: Endogenous Cyclin D1 Promotes the Rate of Onset and Magnitude of Mitogenic Signaling via Akt1 Ser473 Phosphorylation

doi: 10.1016/j.celrep.2020.108151

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: MCF-7 and HEK293T were recently authenticated by ATCC.

Techniques: Virus, Recombinant, In Situ, shRNA, Plasmid Preparation, Mutagenesis, Software