ZFYVE21 Antibody

Western Blot: ZFYVE21 Antibody [H00079038-B01P]

Western Blot: ZFYVE21 Antibody [H00079038-B01P] - Recombinant FLAG-tagged ZFYVE21 (1ug) and/or recombinant SMURF2 (1ug) protein was incubated for 30min at room temperature and analyzed by western blot. Image collected and cropped by CiteAb from the following publication (nature.com/articles/s41467-019-10041-2), licensed under a CC-BY license.

Western Blot: ZFYVE21 Antibody [H00079038-B01P] -

Western Blot: ZFYVE21 Antibody [H00079038-B01P] - Correlations of ZFYVE21 & complement in patient tissues. Western blots of human umbilical vein endothelial cells treated with either panel reactive antibody (30 min, 25% v/v in gelatin veronal buffer) or LIGHT (12 h, 50 ng/mL in EGM2 media, a). Kidney biopsies from renal transplant patients (n = 12) were stained as indicated (b). Synovial biopsies from osteoarthritis (n = 11), low-titer rheumatoid factor antibody rheumatoid arthritis (RF RA; n = 8), & high-titer RF RA (n = 8) were stained as indicated (c–e) & spatial correlations between ZFVYE21 or nuclear factor-inducing kinase (NIK) with endothelial cells (Ulex, f) & membrane attack complex (polyC9, g) were assessed using the computer software (scale bar: 200 μm). ****p < 1 × 10−5, Student’s t test. CD45 abundance was assessed (h) & correlated with NIK staining (i) via blinded scoring from three independent observers. *p < 0.05, **p < 0.001. Western blotting was repeated three times. Analysis of variance statistical analyses were performed. Representative data shown *p < 0.05, **p < 0.001 Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31113953), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ZFYVE21 Antibody [H00079038-B01P] -

Western Blot: ZFYVE21 Antibody [H00079038-B01P] - SMURF2-dependent degradation of vesicular PTEN. PTEN co-immunoprecipitations (co-IPs) were performed on human umbilical vein endothelial cells (HUVECs) treated for 30 min with panel reactive antibody (PRA) sera & blotted as indicated (a). Rab5 co-IPs were performed on PRA-treated HUVECs transfected with control or SMURF2 siRNA (b). Western blot analysis of whole-cell lysates of HUVECs transiently transfected with SMURF2 WT, SMURF2 DN, control siRNA, & SMURF2 siRNA (c). HUVECs transfected with control, SMURF2, or ZFYVE21 siRNA were treated with PRA & Rab5 co-IPs were performed (d). Recombinant FLAG-tagged ZFYVE21 (1 μg) and/or recombinant SMURF2 (1 μg) protein was incubated for 30 min at room temperature & analyzed by western blot (e). In vitro kinase assays were performed using various E2 ubiquitin ligases with SMURF2 protein (1 μg) as the E3 ubiquitin ligase & PTEN (1 μg) as substrate (f). In vitro kinase assays were performed using isolated SMURF2 WT or SMURF2 DN as E3 ubiquitin ligases, UbcH5c as the E2 ubiquitin ligase, & PTEN as substrate as indicated (g). HUVECs were pretreated with MG132 for 4 h or chloroquine for 30 min prior to PRA treatment at the indicated times prior to Rab5 co-IP & western blot analysis (h). Western blot analyses were repeated two to four times. Confocal microscopic studies analyzed ≥4 cells per group & was repeated three times. In vitro kinase assays were repeated twice. Representative data shown Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31113953), licensed under a CC-BY license. Not internally tested by Novus Biologicals.