Journal: Biomaterials

Article Title: Caveolin-Mediated Endocytosis of the Chlamydia M278 Outer Membrane Peptide Encapsulated in Poly(lactic acid)-Poly(ethylene glycol) Nanoparticles by Mouse Primary Dendritic Cells Enhances Specific Immune Effectors Mediated by MHC Class II and CD4+ T cells

doi: 10.1016/j.biomaterials.2017.12.019

Figure Lengend Snippet: DCs were stimulated for 24 hours with PPM, M278 and PPP followed by staining for the subcellular organelles EE (EEA1, early endosome) (A), LE (Rab7, late endosome) (B), ER (endoplasmic reticulum) (C) and lysosome (LAMP-1) (D). Colocalization with organelles was confirmed by probing for the expression of the targeted M278. DAPI (blue) was used to stain the nuclei. The top row (merge) indicate the overlay of images. Direct visualization and imaging were performed employing immunofluorescence microscopy.

Article Snippet: Anti-rab7 (bs-6703R), anti-caveolin-1 (bs-1453R) and anti-LAMP-1(bs-1970R) antibodies were all purchased from Bioss Antibodies Inc. (Woburn, MA).

Techniques: Staining, Expressing, Imaging, Immunofluorescence, Microscopy

Journal: Brain

Article Title: SLC30A9 mutation affecting intracellular zinc homeostasis causes a novel cerebro-renal syndrome

doi: 10.1093/brain/awx013

Figure Lengend Snippet: SLC30A9 protein subcellular localization, effect on Wnt signaling and structural modeling. ( A ) Overexpression of SLC30A9 protein fused to enhanced green fluorescent protein (EGFP) and immunofluorescent staining of different endosomal, Golgi and ER markers (CD63, EEA1, Rab7, GM130 and Calnexin) showing partial co-localization with endoplasmic reticulum in SH-SY5Y cells. On the bottom left side of the merged image of Calnexin marker panel is a magnification of the dashed frame, showing the co-localization in a single cell. Scale bars = 10 µm. ( B ) TOP flash reporter assay of neuroblastoma (SH-SY5Y) cells showing enhanced Wnt signalling in both SLC30A9 wild-type and mutant transfected cells. There was no significant difference between wild-type and mutant transfected cells. ( C ) Structural model of ZnT-9 protein showing the p.(A350del) in the fourth transmembrane helix, putatively causing destabilization of the protein structure in general, affecting the TM metal binding site specifically.

Article Snippet: Cells were then incubated with primary polyclonal goat anti-human EEA1 (Sc-6414; Santa Cruz Biotechnology), polyclonal rabbit anti- human Rab7 (Sc-10767; Santa Cruz Biotechnology), mouse monoclonal anti- human CD63 (CBL553; Merck Millipore) or polyclonal rabbit anti-human Calnexin (Sc-11397; Santa Cruz Biotechnology) antibodies for 1 h. Post-incubation, cells were washed twice with PBST and incubated with secondary donkey anti-goat IgG Alexa Fluor® 546 (A-11056; Invitrogen), goat anti-rabbit IgG Alexa Fluor® 546 (A-11010; Invitrogen) or donkey anti-mouse IgG Alexa Fluor® 546 (A-10036; Invitrogen) accordingly for 1 h. Cells were then washed twice with 1 × PBST and mounted using Vectashield containing DAPI (H-1200; Vector Laboratories).

Techniques: Over Expression, Staining, Marker, Reporter Assay, Mutagenesis, Transfection, Binding Assay