Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: A Bioresponsive Genetically Encoded Antimicrobial Crystal for the Oral Treatment of Helicobacter Pylori Infection.

doi: 10.1002/advs.202301724

Figure Lengend Snippet: Figure 3. Release of GFP from the cleaved Cry3Aa-MIIA(D45E)-GFP crystals. A) Quantitation of GFP signal intensities of MIIACt-GFP released into BB under different scenarios. Fluorescence of the supernatants of each experimental sample from the GFP release assay (upper) and their corresponding GFP signal intensities as measured using a TECAN (lower). Data are represented as mean ± SEM (N = 3). ****P < 0.0001. ns, not significant. B) Release of MIIACt-GFP from cleaved crystals under different conditions was detected by western blot using anti-GFP antibody. C) Crystal suspensions before and after incubation for 48 h in different buffers. Significant degradation of the released MIIACt-GFP in the no Ca2+ pH 7.5 buffer was observed. D) Corresponding release profiles of the MIIACt-GFP fragment in (B). Data are represented as mean ± SEM (N = 2). E) SDS-PAGE analysis of the binding of wild-type MIIA protein to Cry3Aa crystals in the presence of 15 mm CaCl2. Lanes 2–4 are protein controls. Lanes 5 and 6 are the Cry3Aa-bound MIIA complex pellet (P) and supernatant (SN) after centrifugation. F,G) SDS-PAGE analyses and corresponding bar plots on the release of MIIACt from Cry3Aa-bound MIIA complex in F) pH 4.0 or pH 7.5 PBS, and G) in 15 mm CaCl2 or no CaCl2, pH 7.5 buffer. P: pellet, SN: supernatant. H) Efficient binding of MIIA(D45A) protein to Cry3Aa crystals in the presence of CaCl2. I) Schematic illustration of the Ca2+-medicated cleavage and binding of MIIACt-GFP. Subfigures (E) and (F) are part of the same SDS-PAGE gel sharing one molecular weight marker. The original gel image is provided in Figure S8 (Supporting Information). Subfigures (G) and (H) are derived from the same SDS-PAGE gel and thus share one marker lane. The original gel image is provided in Figure S9 (Supporting Information). Subfigures (F) and (H) were produced by splicing the marker lane and the relevant lanes of the original gel image.

Article Snippet: The MIIACt-GFP released into the supernatant was detected using anti-GFP (rabbit) primary antibody (Rockland 600-401-215, 1:2000) and HRP conjugated secondary antibody of goat anti-rabbit IgG (Bio-Rad 170–6515, 1:5000).

Techniques: Quantitation Assay, Fluorescence, Release Assay, Western Blot, Incubation, SDS Page, Binding Assay, Centrifugation, Molecular Weight, Marker, Derivative Assay, Produced

Journal: BMC Research Notes

Article Title: Melanocortin agonists stimulate lipolysis in human adipose tissue explants but not in adipocytes

doi: 10.1186/s13104-015-1539-4

Figure Lengend Snippet: Non-esterified fatty acid (NEFA) release in human subcutaneous adipocytes: a Adipocytes purchased from ZenBio were incubated with increasing concentrations (1 pM–10 μM) of positive control isoproterenol (EC 50 = 1.46 ± 0.34 nM), ACTH, α-MSH, PG-901 and LY2112688 (n = 4). NEFA release was measured as described. Experiments were carried out in duplicates. Data are shown as average values ± SEM. b MC1-5R and GAPDH mRNA expression was measured in same adipocytes (n = 3 pooled). cDNA was synthesized and mRNA levels determined by standard PCR (cDNA 1:10 from 1 μg RNA, 45 cycles)

Article Snippet: Fig. 2 Non-esterified fatty acid (NEFA) release in human subcutaneous adipocytes: a Adipocytes purchased from ZenBio were incubated with increasing concentrations (1 pM–10 μM) of positive control isoproterenol (EC 50 = 1.46 ± 0.34 nM), ACTH, α-MSH, PG-901 and LY2112688 (n = 4).

Techniques: Incubation, Positive Control, Expressing, Synthesized