Journal: Kidney international

Article Title: The L1 cell adhesion molecule is a potential biomarker of human distal nephron injury in acute tubular necrosis.

doi: 10.1038/sj.ki.5002640

Figure Lengend Snippet: Figure 4 | Comparative localization of L1 intra- and extracytoplasmic domains. (a) L1 scheme showing the extracellular part with six immunoglobulin-like domains (Ig-like) (including one recognized by mAb272) and five fibronectin III-like (FNIII) domains followed by the transmembrane segment and cytoplasmic fragments (recognized by the goat polyclonal serum). (b–d) Normal CDs are compared with (e–g) damaged CDs. (b, e) CDs are stained by goat polyclonal antiserum labeled by FITC (green) and (c, f) mAb272 labeled by rhodamine (red). (d, g) Merge. (d) In normal CD, both antibodies give a basolateral signal. (g) In injured principal cells, the basolateral signal is low with both antibodies, whereas there is a strong apical membrane (arrow) signal with the anti-cytoplasmic domain antibody contrasting with no signal for the extracellular fragment. Original magnification 630.

Article Snippet: After four washes with TBS-T (Tris-buffered saline with 0.1% Tween), goat anti-human L1 extracellular domain antibody was added (R&D Systems), followed by anti-goat coupled to alkaline phosphatase (Sigma-Aldrich, St Louis, MO, USA).

Techniques: Staining, Labeling, Membrane

Journal: bioRxiv

Article Title: Strain-Specific Human Natural Killer Cell Recognition of Influenza A Virus

doi: 10.1101/148528

Figure Lengend Snippet: (A) Cytokine concentrations assessed by Luminex®, values displayed represent the mean fluorescence intensity in pH1N1-infected/H3N2 infected conditions (MOI=3). Cytokines elevated by 2.5-fold over the level in mock-infected monocytes are plotted. (B) Impact of cytokine receptor blocking on the NK cell IFN-γ response evaluated by pre-incubating NK cells for 1 hr with blocking antibodies specific to IFNAR2, IL-12R, IL-15R, IL-18R, and IFNGR1 followed by co-culture with infected monocytes. 24 HPI, intracellular cytokine staining was used to assess NK cell IFN-γ + frequency compared to treatment with an isotype control antibody (H3N2: n =2-6, pH1N1: n =9). (C) NK cells were incubated for 1 hr with antibodies specific to IFNAR2 and CD226 or CD54 followed by co-culture with autologous infected monocytes. At 24 HPI, intracellular cytokine staining was used to assess IFN-γ production compared to treatment with an isotype control antibody ( n =6). * P < 0.05, ** P < 0.005, Wilcoxon signed-rank test. (D) Model of strain-specific NK cell recognition of influenza A infection. pH1N1-infection of monocytes virus does not downregulate CD54 and CD112 to the same extent as H3N2-infected monocytes. CD54 expression is retained on Flu-NP + cells while CD112 expression is preferentially retained on exposed, uninfected monocytes. pH1N1 infection of monocytes elicits enhanced IFN-α secretion and blockade of IFNAR2 dampens NK cell anti-pH1N1 IFN-γ production.

Article Snippet: IL-12R β 2 Ab (R&D Systems AF1959-SP) was used at a final concentration of 83.33 μg/mL.

Techniques: Luminex, Fluorescence, Infection, Blocking Assay, Co-Culture Assay, Staining, Control, Incubation, Virus, Expressing

Journal: Journal of Microbiology and Biotechnology

Article Title: Inhibitory Effects of Latilactobacillus curvatus BYB3 Cell-Free Extract on Human Melanoma B16F10 Cells and Tumorigenic Mice

doi: 10.4014/jmb.2309.09002

Figure Lengend Snippet: ( A ) The cells were first stained with crystal violet solution. Subsequent examination showed that the three strain extracts were not affected by the malignancy in the B16F10 cells; however, the proliferation of BYB3-treated cell was inhibited more than that of LGG and GP1B treated cells. ( B ) Viability began to decrease in a dose-dependent manner in the B16F10 cells as the concentration of the three strain extracts decreased from 75 μg/ml to lower concentrations than in BYB3. Statistically significant differences are shown. (* p < 0.05, ** p < 0.01).

Article Snippet: The member was blocked with 5% skim milk (Difco Laboratories) and underwent overnight antibody incubation Primary Abs against the following proteins were used: human/mouse NLRP3 (1:1000, AP-20B-0014, AdipoGen), human NCOA6 (1:1000, A300-411A, Bethyl Laboratories), human/mouse ASC (1:1000, sc-514414, Santa Cruz Biotechnology; AG-25B-0006, AdipoGen), IL-1β (1:500, AF-401-NA, R&D Systems), mouse cleaved caspase-1 (1:1000, AG-20B-0042, AdipoGen), FLAG (1:1000, sc-166355, Santa Cruz Biotechnology), Myc (1:1000, ab13836, Abcam), GAPDH (1:1000, sc-25778, Sigma).

Techniques: Staining, Concentration Assay