EB1 (MAPRE2, microtubule-associated protein, RP/EB family, member 2, EB2, RP1) may influence tumorigenesis of colorectal cancers and proliferative control of normal cells. EB1 may belong to the intermediate/early gene family, involved in the signal transduction cascade downstream of the TCR. Colorectal cancer is caused by the pathologic transformation of normal colonic epithelium to an adenomatous polyp, which can become an invasive cancer. APC (adenomatous polyposis coli) is a tumor suppressor gene, the mutation of which is one of the earliest events in colorectal carcinogenesis. A majority of the mutations result in the loss of the carboxy terminus of APC. EB1 has been shown to bind to the carboxy terminal region of APC, which implicates EB1 in APC suppression of colonic cancer. EB1 overexpression may play a role in the development of human esophageal squamous cell carcinoma (ESCC) by affecting APC function and activating the beta-catenin/TCF pathway. EB3 is related to EB1 and likewise associates with the microtubule cytoskeleton. EB3 is expressed predominantly in the central nervous system and preferentially associates with APCL.
Background References
1. Yang C et al. EB1 and EB3 regulate microtubule minus end organization and Golgi morphology. J Cell Biol 216:3179-3198 (2017).
2. Komarova Y et al. Mammalian end binding proteins control persistent microtubule growth. J Cell Biol 184:691-706 (2009).
Western blot analysis of EB3 on different lysates with Rabbit anti-EB3 antibody (ET7106-99) at 1/2,000 dilution.
Lane 1: SH-SY5Y cell lysate (20 µg/Lane) Lane 2: U-87 MG cell lysate (20 µg/Lane) Lane 3: MG-63 cell lysate (20 µg/Lane) Lane 4: Mouse brain tissue lysate (20 µg/Lane) Lane 5: Rat brain tissue lysate (20 µg/Lane)
Predicted band size: 32 kDa Observed band size: 35 kDa Exposure time: Lane 1-3: 30 seconds; Lane 4-5: 8 seconds; ECL: K1801; 4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7106-99) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
EB3 was immunoprecipitated from 0.2 mg U-87 MG cell lysate with ET7106-99 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET7106-99 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: U-87 MG cell lysate (input) Lane 2: ET7106-99 IP in U-87 MG cell lysate Lane 3: Rabbit IgG instead of ET7106-99 in U-87 MG cell lysate