Journal: PLoS ONE

Article Title: N-Glycosylation of the Na+-Taurocholate Cotransporting Polypeptide (NTCP) Determines Its Trafficking and Stability and Is Required for Hepatitis B Virus Infection

doi: 10.1371/journal.pone.0170419

Figure Lengend Snippet: Effect of N-linked glycosylation on NTCP protein expression. (A and B) HA-tagged NTCP was immunoprecipitated (IP) from lysates of HepG2 cells stably expressing NTCP-WT, NTCP-N5Q, NTCP-N11Q or NTCP-N5,11Q. IP samples were subjected to immunoblotting for NTCP (using anti-HA-hrp). (B) IP samples were digested with PNGase F for 1h at 37°C (500 units) prior to immunoblotting for NTCP. (C) NTCP was quantified by image J and expressed relative to NTCP-WT. Molecular mass is given in kDa on the left-hand side. Results are mean +/- sd. *Indicates significant different from NTCP-WT (p

Article Snippet: Samples were analyzed by immunoblotting using anti-HA antibodies (Sigma) and HRP-conjugated mouse antibodies (Sigma) as described above.

Techniques: Expressing, Immunoprecipitation, Stable Transfection

Journal: BioMed Research International

Article Title: Induction of Boosted Immune Response in Mice by Leptospiral Surface Proteins Expressed in Fusion with DnaK

doi: 10.1155/2014/564285

Figure Lengend Snippet: Analysis of IgG production induced in mice by recombinant proteins. The sera from mice immunized with fused-proteins were analyzed by ELISA (a) and Western blotting (b). In (a) wells were coated with DnaK or surface proteins, as depicted; in (b) blotted proteins are DnaK (lane 1), Lsa21 (lane 2), rLIC10494 (lane 3), Lp95 C-terminus region (lane 4), and rLIC12730 (lane 5). M: molecular mass protein marker. In both methods, proteins were probed with the respective antifusion protein serum (1 : 20,000 dilution) and the reactions were developed with HRP-conjugated anti-goat IgG (1 : 10,000) (ELISA) and HRP-conjugated anti-mouse IgG (1 : 5,000) (Western).

Article Snippet: After washing, the membrane was incubated with HRP-conjugated anti-mouse IgG (1 : 5,000; Sigma) in 5% nonfat dried milk/PBS-T for 1 h. The bands were revealed with ECL reagent kit chemiluminescence substrate (GE Healthcare).

Techniques: Mouse Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Western Blot, Marker

Journal: The Journal of Cell Biology

Article Title: The Ig cell adhesion molecule Basigin controls compartmentalization and vesicle release at Drosophila melanogaster synapses

doi: 10.1083/jcb.200701111

Figure Lengend Snippet: Bsg accumulation at the NMJ. (A and B) NMJs of w (A) or protein-trap/+ (B) larvae double stained with anti-Bsg (A) or anti-GFP (B) antibodies and anti-Dlg antibodies (A′ and B′). (C) elav -Gal4/+; UAS-GFP- bsg /+ larvae stained with anti-GFP (C) and anti-HRP (C′) antibodies. (D and E) NMJs of w (D) and protein-trap/+ (E) larvae double stained with anti-Bsg (D) or anti-GFP (E) and anti-HRP (D′ and E′) antibodies. A–E correspond to z projections of serial confocal sections. Bar, 10 μm. (F) Tangential confocal section of a protein-trap/+ synaptic bouton stained with anti-GFP and NC82 anti-BRP antibodies and plot of the intensity profiles of GFP (green) and NC82 (pink) stainings across the bouton (section is indicated by white marks on the overlay). A–E are from muscle 6/7 NMJs, and F is from muscle 4 NMJ. Bar, 5 μm.

Article Snippet: The following antibodies were used for Western blot analysis: rabbit anti-GFP (1:500 [Santa Cruz Biotechnology, Inc.] or 1:2,500 [Torrey Pines]) and HRP-conjugated anti–mouse, –rat, or –rabbit (1:2,000; GE Healthcare) antibodies.

Techniques: Staining

Journal: The Journal of Cell Biology

Article Title: The Ig cell adhesion molecule Basigin controls compartmentalization and vesicle release at Drosophila melanogaster synapses

doi: 10.1083/jcb.200701111

Figure Lengend Snippet: Distribution of actin cytoskeleton markers is altered in bsg larvae. (A and B) Wild-type (A) and Df/bsg 6293 (B) muscle 4 NMJs stained with anti–α-Spec antibodies. Bar, 10 μm. (C–E) Heterozygous control (C) and Df/bsg 6293 (D and E) boutons stained with anti–α-Spec (C1, D1, and E1), anti-HRP (C2, D2, and E2), and anti-Dlg (C4, D4, and E4) antibodies. Bar, 5 μm. (F and G) w (F) and Df/bsg 1217 (G) bouton stained with anti-Wasp (F and G) and anti–α-Spec (F′ and G′, red) antibodies. (H and I) w (H) and Df /bsg 6293 (I) boutons stained with anti–α-Spec (H and I) and anti-Fusch (H′ and I′) antibodies. Bar, 5 μm. (J and K) Synaptic boutons of wild-type (J) and Df/bsg 6293 (K) larvae expressing a fusion of GFP with the F-actin binding domain of Moesin (GMA), under the control of elav -Gal4. GFP-GMA expression is shown in J and K, and is in green in J′ and K′. HRP staining is shown in red in J′ and K′. Bar, 5 μm. Images A and B correspond to z projections of serial confocal sections throughout entire boutons (step size: 0.3 μm), and images C–K correspond to single optical slices taken through bouton centers. (L) Graph showing the percentage of NMJ 6/7 branches containing presynaptic Spec + or GMA + aggregates. **, P

Article Snippet: The following antibodies were used for Western blot analysis: rabbit anti-GFP (1:500 [Santa Cruz Biotechnology, Inc.] or 1:2,500 [Torrey Pines]) and HRP-conjugated anti–mouse, –rat, or –rabbit (1:2,000; GE Healthcare) antibodies.

Techniques: Staining, Expressing, Binding Assay

Journal: The Journal of Cell Biology

Article Title: The Ig cell adhesion molecule Basigin controls compartmentalization and vesicle release at Drosophila melanogaster synapses

doi: 10.1083/jcb.200701111

Figure Lengend Snippet: GFP expression pattern in bsg protein-trap lines. (A and A′) Third instar larva heterozygous for the protein-trap insertion, stained with anti-GFP antibodies (A and A′, green), anti-HRP antibodies (A′, red), and phalloidin (A′, blue). (B) GFP-Bsg accumulation in heterozygous larval brain. (C) Western blot of total extracts from w larvae (left) or larvae heterozygous (protein-trap/+; middle) or homozygous (protein-trap; right) for a protein-trap insertion, probed with anti-Bsg and anti-GFP antibodies. (D) Alignment of D. melanogaster and human Bsg proteins. The green triangle indicates the location of GFP insertion.

Article Snippet: The following antibodies were used for Western blot analysis: rabbit anti-GFP (1:500 [Santa Cruz Biotechnology, Inc.] or 1:2,500 [Torrey Pines]) and HRP-conjugated anti–mouse, –rat, or –rabbit (1:2,000; GE Healthcare) antibodies.

Techniques: Expressing, Staining, Western Blot