Journal: PLoS ONE

Article Title: Gain in Brain Immunity in the Oldest-Old Differentiates Cognitively Normal from Demented Individuals

doi: 10.1371/journal.pone.0007642

Figure Lengend Snippet: Significantly affected genes ( p <0.05) in the oldest-old (≥87 y.o.) cohort in comparison to cognitively intact youngest-old (≤86 y.o.) subjects.

Article Snippet: Blots were incubated with antibodies: rabbit anti-human CX3C chemokine fractalkine receptor 1, CX3CR1 (1∶500 v/v) from LifeSpan Biosciences Inc. (Seattle, WA); mouse anti-human HLA-DPA1 (1∶100 v/v) from Abnova (Walnut, CA); mouse mAb against human CD74 antigen protein (1∶1000 v/v), which recognizes cytoplasmic tail of CD74 (Cedarlane/StressMarq, Burlington, NC) and mouse anti-human β tubulin (TUBB; 1∶1000 v/v) – from Santa Cruz Biotech.

Techniques: Binding Assay

Journal: PLoS ONE

Article Title: Gain in Brain Immunity in the Oldest-Old Differentiates Cognitively Normal from Demented Individuals

doi: 10.1371/journal.pone.0007642

Figure Lengend Snippet: Expression analysis of the immune response genes in the inferior temporal cortex - BA20.

Article Snippet: Blots were incubated with antibodies: rabbit anti-human CX3C chemokine fractalkine receptor 1, CX3CR1 (1∶500 v/v) from LifeSpan Biosciences Inc. (Seattle, WA); mouse anti-human HLA-DPA1 (1∶100 v/v) from Abnova (Walnut, CA); mouse mAb against human CD74 antigen protein (1∶1000 v/v), which recognizes cytoplasmic tail of CD74 (Cedarlane/StressMarq, Burlington, NC) and mouse anti-human β tubulin (TUBB; 1∶1000 v/v) – from Santa Cruz Biotech.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Gain in Brain Immunity in the Oldest-Old Differentiates Cognitively Normal from Demented Individuals

doi: 10.1371/journal.pone.0007642

Figure Lengend Snippet: Protein levels of CD74 were measured by Western Blot . Data expressed as means ± SEM of ODs normalized to the generic sample – “calibrator” and endogenous control - TUBB protein (N = 9–10/group, see ).

Article Snippet: Blots were incubated with antibodies: rabbit anti-human CX3C chemokine fractalkine receptor 1, CX3CR1 (1∶500 v/v) from LifeSpan Biosciences Inc. (Seattle, WA); mouse anti-human HLA-DPA1 (1∶100 v/v) from Abnova (Walnut, CA); mouse mAb against human CD74 antigen protein (1∶1000 v/v), which recognizes cytoplasmic tail of CD74 (Cedarlane/StressMarq, Burlington, NC) and mouse anti-human β tubulin (TUBB; 1∶1000 v/v) – from Santa Cruz Biotech.

Techniques: Western Blot

Journal: Frontiers in Immunology

Article Title: Faecalibacterium prausnitzii Skews Human DC to Prime IL10-Producing T Cells Through TLR2/6/JNK Signaling and IL-10, IL-27, CD39, and IDO-1 Induction

doi: 10.3389/fimmu.2019.00143

Figure Lengend Snippet: DC exposure to F. prausnitzii switches their cytokine profile from pro-inflammatory to anti-inflammatory. IL-12p70 (A) , TNF-a (B) , and IL-10 levels (C) secreted in response to LPS by DC exposed or not (Ctrl) to Fprau at the beginning of their differentiation. (D) Representative dot plot of intracellular IL-12p35 and p40 co-labeling in LPS-stimulated DC exposed or not to F. prausnitzii . (E) LPS-induced IL-12p40 ( n = 6–24) and IL-12p35 ( n = 9–27) expression by DC exposed or not (Ctrl) to indicated bacteria (ratio:1:1) during the last 48 h. (F) IL-12p40 ( n = 12) and p35 ( n = 6) expression by myeloid DC maintained for 24 h in culture with or without F. prausnitzii , and then stimulated 12 h with LPS. Wilcoxon test. * p < 0.05, ** p < 0.005, *** p < 0.0005, **** p < 0.00005.

Article Snippet: We used: PE-labeled antibodies to CD40 (clone 5C3), CD80 (clone L307.4), CD83 (clone HB15e), CD39 (clone TU66), PDL-1 (clone MIH1), as well as anti-CD4-APC (clone SK3) all from Becton Dickinson: (BD), anti-CD86-PE (clone HA5.2B7, Beckman Coulter), anti-IL-12p35/p70-PE (clone REA121, Miltenyi Biotech), anti-IL-12/IL-23p40-APC (clone C11.5, BioLegend), anti-IL-10-PE (clone JES3-19F1, BD), anti-IFN-γ-APC (clone B27, BD), anti-IL-13-PE (clone JES10-5A2, BD), anti-IDO-1-APC (clone 70083, R&D), anti-HO-1 (HO-1-1, Thermoscientific).

Techniques: Labeling, Expressing, Bacteria

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Monophosphoryl lipid A plus IFNγ maturation of dendritic cells induces antigen-specific CD8 + cytotoxic T cells with high cytolytic potential

doi: 10.1007/s00262-010-0843-z

Figure Lengend Snippet: The superior induction of CTLs by MPLA/IFNγ DCs compared to CC-DCs is dependent on IL-12 and not on IL-6 or IL-23. a Blocking antibodies specific for IL-12p40 and IL-6, and an irrelevant antibody were added to MPLA/IFNγ DC-CTL cocultures. 7 days after the second stimulation of the CTLs with DCs (day 17), the CTLs were stained with the MART-1 tetramer (TM) and CD8-FITC. A graph showing the % MART-1 TM cells for all separate cultures of one experiment is shown. The condition treated with anti-IL-12 significantly differed (one-way ANOVA, P < 0.05) from untreated or isotype control treated conditions. A representative experiment out of four is depicted. b Blocking antibodies specific for IL-12p35 and IL-23p19, and an irrelevant antibody were added to MPLA/IFNγ DC-CTL cocultures. 7 days after the third stimulation of the CTLs, the CTLs were stained with the MART-1 TM and CD8-FITC. A graph showing the % MART-1 TM cells for all separate cultures of one experiment is shown. The condition treated with anti-IL-12p35 significantly differed (one-way ANOVA, P < 0.01) from isotype control treated condition

Article Snippet: The following neutralizing antibodies were used; anti-IL-12p40 (C8.6), anti-IL-12p35 (B-T21, Diaclone), anti-IL-23p19 (B-Z23, Diaclone), anti-IL-6 (IL6.8 [ 26 ]).

Techniques: Blocking Assay, Staining, Control

Journal: Scientific Reports

Article Title: Defective membrane repair machinery impairs survival of invasive cancer cells

doi: 10.1038/s41598-020-77902-5

Figure Lengend Snippet: Expression and subcellular localization of endogenous Anx in MDA-MB-231. ( a ) 10 µg of protein extracts from MDA-MB-231 and BeWo cells were analyzed by western-blot. The gel analysis plugin of ImageJ was used to quantify the density of bands. The histogram presents mean values (± SD) of the ratio Anx/GAPDH from ten independent experiments. Student t Test for independent samples. ** p < 0.01. ( b ) Revelation of AnxA5 in MDA-MB-231 and BeWo cells compared to 50 ng purified recombinant AnxA5 (Pure A5), presented as an example. The whole membrane, together with representative western-blot analysis for AnxA1, A2, A4 and A6, are presented in Supplementary Fig. . ( c ) MDA-MB-231 cells were immunostained for AnxA1, AnxA2, AnxA4, AnxA5 and AnxA6 (green), as indicated, and counterstained with DAPI (blue). The right-hand column presents magnified images extracted from the “Anx” column (indicated by inserts). Scale bar: 40 µm. ( d ) Quantification of fluorescence intensities after immunostaining of endogenous Anx in MDA-MB-231 cells. Fluorescence intensities were measured from 8-bit images by drawing a circular ROI inside the cytoplasm and measuring the mean pixel value. Mean pixel values from at least 20 cells are presented. Horizontal bars represent median values. The variance value is 323, 268, 224, 2372 and 2909 for AnxA1, A2, A4, A5 and A6, respectively. The coefficient of variation is 14%, 13%, 24%, 35% and 47% for AnxA1, A2, A4, A5 and A6, respectively. The expression level of AnxA5 or AnxA6 is heterogenous from one MDA-MB-231 cell to another. In some cells, AnxA5 and AnxA6 are expressed at a very low level.

Article Snippet: Semi-dry electrophoretic transfer (Bio-Rad, Hercules, CA, USA) onto PVDF membrane was performed for 1 h at 100 V. The cellular content in AnxA1 (37 kDa), AnxA2 (36 kDa), AnxA4 (32 kDa), AnxA5 (35 kDa), AnxA6 (68 kDa), and glyceraldehyde-3-phosphate deshydrogenase (GAPDH, loading control, 37 kDa) was detected with rabbit anti-AnxA1 polyclonal antibody (PA1006, BosterBio, Pleasanton, CA, USA), mouse anti-AnxA2 monoclonal antibody (3E8-B6, Sigma, Saint-Louis, MO, USA), mouse anti-AnxA4 monoclonal antibody (SAB4200121, Sigma), mouse anti-AnxA5 monoclonal antibody (AN5, Sigma), mouse anti-AnxA6 monoclonal antibody (sc-271859, Santa cruz Biotechnology), and rabbit anti-GAPDH polyclonal antibody (FL-335, Santa Cruz Biotechnology), respectively.

Techniques: Expressing, Western Blot, Purification, Recombinant, Membrane, Fluorescence, Immunostaining