anti-arf6 ab  (ZenBio)

Journal: Journal of Nanobiotechnology

Article Title: GRP75-driven, cell-cycle-dependent macropinocytosis of Tat/pDNA-Ca 2+ nanoparticles underlies distinct gene therapy effect in ovarian cancer

doi: 10.1186/s12951-022-01530-6

Figure Lengend Snippet: Cell-cycle-dependent centrosomal enrichment of GRP75 and its phosphorylation acting on sub-phase distribution. (A) Cyclebase-based predication of expression of GRP75, MPS1, VDAC1 and Arf6 in sub-phases of cell-cycle (dark blue content indexes the mRNA level during cell-cycle). (B) Western blot determined the expression level of those mentioned protein in subcellular fractions of synchronized Skov3. Protein bands were quantified by Image J software and expression ratio (compared to cells without drug-induction, Ctrl, set as 1) in the sub-phase are correspondingly marked below. (C) Schematic GRP75 constructs: wt: wild type. unphospho: unphosphorlation (T62A/S65A). phospho mimic: phosphorlation mimic (T62D/S65D). (D ) Lentivirus-stable-transduced Skov3 and Cos7 cells were built based on GRP75-targeting shRNA (GRP75-KD), or over-expression (OE) with constructs as listed above. GRP75 expression level was analyzed by western blotting. ( E ) Bar graphs show the sub-phase percentage of cell-cycle in GRP75-KD or -OE cells. Data are the average results of three independent experiments

Article Snippet: Rabbit anti-Arf6 Ab (381658) was obtained from Zenbio.

Techniques: Phospho-proteomics, Expressing, Western Blot, Software, Construct, shRNA, Over Expression

Journal: PLoS ONE

Article Title: TNF Stimulates Nuclear Export and Secretion of IL-15 by Acting on CRM1 and ARF6

doi: 10.1371/journal.pone.0069356

Figure Lengend Snippet: (A) ARF6 was bound to IL-15Rα. Cells were transfected with ARF6-HA 24 h earlier. ARF6 protein was immunoprecipitated and subjected to WB for IL-15 and IL-15Rα. Immunoprecipitation with nonspecific IgGs was done as the control. (B) ARF6 co-localized with IL-15Rα in the plasma membrane and some organelles. After co-transfection, ARF6-HA (green) and IL-15Rα (red) were co-localized, as shown by confocal microscopy. Bar: 10 μm.

Article Snippet: Goat anti IL-15 antibody (Ab) (SC-1296 ), goat anti-IL-15Rα Ab (SC-1524), rabbit anti-IL-15 Ab (SC-788900), rabbit anti-IL-15Rα (SC-9172) Ab, rabbit anti-calnexin Ab (SC-6465) and goat anti Rab7 Ab (SC-6563), were purchased from Santa Cruz Biotechnology (Santa Cruz, CA); Rabbit-β-COP Ab and rabbit-EEA Ab were obtained from ABR (Golden, CO); Rabbit anti-Arf6 Ab was purchased from Proteintech Group (Chicago, IL).

Techniques: Transfection, Immunoprecipitation, Control, Clinical Proteomics, Membrane, Cotransfection, Confocal Microscopy

Journal: PLoS ONE

Article Title: TNF Stimulates Nuclear Export and Secretion of IL-15 by Acting on CRM1 and ARF6

doi: 10.1371/journal.pone.0069356

Figure Lengend Snippet: HeLa cells were transfected with pCDNA3.1 (control), WT-ARF6, or DN-ARF6 plasmids 48 h before the assay. (A) In control cells and those overexpressing WT-ARF6, TNF treatment for 4 h increased IL-15 release. The cells overexpressing DN-ARF6 did not show a response. In none of the groups did PBS vehicle treatment increase IL-15 release. Data are representative of three independent experiments. *: p<0.05 compared with the control. (B) The overexpression of WT-ARF6 and/or TNF treatment seemed to increase the expression of IL-15 and IL-15Rα, shown by WB of whole cell lysates. (C) The potential interaction of ARF6 with IL-15 was shown by ICC. The left panel is ARF6 immunoreactivity (red, stained with anti-ARF6 in control cells and anti-HA in cells overexpressing ARF6) and the right panel is IL-15 immunoreactivity (green). In control cells, endogenous ARF6 showed a diffuse vesicular pattern of cytoplasmic distribution. In cells overexpressing WT-ARF6 that increased overall expression as well as cell surface distribution of ARF6, IL-15 signal was also increased and showed cell surface distribution co-localizing with WT-ARF6. In cells overexpressing DN-ARF6, the small increase of ARF6 signal remained cytoplasmic, and did not show co-localization with IL-15. Bar: 10 μm. D) Intracellular distribution of IL-15 Rα (green) did not show co-localization with endogenous ARF6 in control cells (stained with an ARF6 Ab, red) or in those overexpressing WT-ARF6 (middle panel) or DN-ARF6 (lower panel, HA Ab staining, red). Bar: 10 μm.

Article Snippet: Goat anti IL-15 antibody (Ab) (SC-1296 ), goat anti-IL-15Rα Ab (SC-1524), rabbit anti-IL-15 Ab (SC-788900), rabbit anti-IL-15Rα (SC-9172) Ab, rabbit anti-calnexin Ab (SC-6465) and goat anti Rab7 Ab (SC-6563), were purchased from Santa Cruz Biotechnology (Santa Cruz, CA); Rabbit-β-COP Ab and rabbit-EEA Ab were obtained from ABR (Golden, CO); Rabbit anti-Arf6 Ab was purchased from Proteintech Group (Chicago, IL).

Techniques: Transfection, Control, Over Expression, Expressing, Staining