Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Rheumatoid arthritis synovial stromal cells inhibit apoptosis and up-regulate Bcl-xL expression by B cells in a CD49/CD29-CD106-dependent mechanism.

doi: 10.4049/jimmunol.164.2.1110

Figure Lengend Snippet: FIGURE 7. Stimulation by VLA-4 can induce Bcl-xL protein and mRNA in B cells. B cells (1 3 106) were cultured for 3 days with mAbs to CD11a/CD18 and/or CD29/CD49d in 24-well culture plates coated with rabbit anti-mouse IgG. Afterward, the expression of Bcl-2 and Bcl-xL pro- tein was assessed by Western blotting, and the percentages of viable and apoptotic cells were measured as described in Materials and Methods, and mRNA for Bcl-2, Bcl-xL, and G6PD were assessed by RT-PCR as de- scribed in Materials and Methods. The results of Southern blotting are shown in Expt. 1, and the expression of PCR products by ethidium bromide staining is shown in Expt. 2. One to three micrograms of cDNA was used for each amplification. The number of PCR cycles was modified to ensure that the PCR products obtained were from the linear phase of amplification. The cycle number of each PCR is shown in the figure. Representative data from two of five experiments with similar results are shown.

Article Snippet: Mouse IgG1 (MOPC) mAb, mouse anti-human IgM heavy chain (DA4.4) conjugated with biotin, mouse anti-human CD11a mAb (TS1/22), and mouse antihuman CD18 mAb (TS1/18) were prepared from hybridoma cell lines purchased from American Type Culture Collection (Manassas, VA).

Techniques: Cell Culture, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Southern Blot, Staining

Journal: Channels

Article Title: Cone dystrophy and ectopic synaptogenesis in a Cacna1f loss of function model of congenital stationary night blindness (CSNB2A)

doi: 10.1080/19336950.2017.1401688

Figure Lengend Snippet: Primary antibodies used in this study.

Article Snippet: PKCα , Mouse , Purified Mouse Monoclonal IgG2a , 1:1000 , Cedarlane Labs, Burlington, ON , purified bovine brain PKCα (Clone MC5) , [ ] .

Techniques: Affinity Purification, CRAfT Assay, Purification, Recombinant

Journal: Frontiers in Immunology

Article Title: CCR7 Is Recruited to the Immunological Synapse, Acts as Co-stimulatory Molecule and Drives LFA-1 Clustering for Efficient T Cell Adhesion Through ZAP70

doi: 10.3389/fimmu.2018.03115

Figure Lengend Snippet: ZAP70 controls CCR7-mediated inside-out signaling to LFA-1. (A) Arrest of Jurkat P116 cells lacking ZAP70 and Jurkat P116 cells reconstituted with ZAP70-GFP on immobilized ICAM-1 in the presence or absence of CCL19, CCL21, or MgCl 2 . Mean ± SEM of three independent experiments. (B) Binding of the β 2 -integrin high-affinity reporter mAb24 to Jurkat P116 and Jurkat P116 ZAP70-GFP cells upon CCR7 triggering for 10 min. Mean ± SEM of five independent experiments. (C–E) Confocal images of CD11a staining on Jurkat P116, Jurkat P116 ZAP70-GFP, or Jurkat P116 ZAP70-K369R-GFP cells before and after chemokine stimulation (0.5 μg/ml, 10 min). (C) no pre-treatment. (D) pre-treatment with DMSO or piceatannol. Representative images derived from one out of three independent experiments; scale bar, 10 μm. **** p < 0.0001. ns, not significant.

Article Snippet: The following antibodies were used: PE-labeled anti CD69 (clone FN50) (Bio-Rad, Hercules, CA, USA), PE-labeled anti-IL-2 (BD Pharmigen, Franklin Lakes, NJ, USA), anti-human CCR7 used for immune fluorescence (SAB4500329) (Sigma-Aldrich, St. Louis, MO, USA), anti-human CCR7 (LifeSpan Biosciences, Seattle, WA, USA) used for PLA, anti-human Vav1 (9C1) (Abnova, Taipei City, Taiwan) used for PLA, anti-human CCR7 APC (FAB197A) (R&D Systems, Minneapolis, MN, USA), anti-humanCD3ζ (ab188850) (abcam, Cambridge, United Kingdom), anti-CD3 (ab5690) (abcam), anti-CD3 (clone OKT3; Janssen-Cilag, Beerse, Belgium), anti-ZAP70 (D1C10E) XP® Rabbit mAb (Cell Signaling, Danvers, MA), anti-ZAP70 (phospho Y319) antibody (ab131270) (abcam), anti-YFP1 (E385) (abcam), anti-YFP2 (11814460001) (Roche, Basel, Switzerland), monoclonal anti-HA-HRP (clone HA7) (Sigma-Aldrich), anti-HA antibody (clone HA7) (Sigma-Aldrich), anti-CD11a antibody [EP1285Y] (ab52895) (abcam), anti-CD18 antibody [MEM-48] (ab657) (abcam), mAb24 anti-CD11+CD18 antibody [24] (ab13219) (abcam).

Techniques: Binding Assay, Staining, Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: Jurkat T-Cell Antigen-Independent Elimination of PMA-Activated Neuroblastoma Cells Is Triggered by CCL2/CCR2, Depends Upon Lipid Raft LFA1/ICAM1 Immune Synapses, Is Mediated by m-TRAIL and Is Augmented by the TrkAIII Oncoprotein

doi: 10.3390/ijms27041970

Figure Lengend Snippet: ( a ) Representative indirect IF micrographs and Western blots demonstrating constitutive immunoreactivity and expression of Jurkat CD11a and ICAM-1 (red) and TrkAIII-SH-SY5Y ICAM-1 (red), which is not increased by PMA treatment (60 ng/mL for 16 h), nuclei are stained blue with DAPI (bar = 10 μm). ( b ) Representative negative-image micrographs demonstrating Jurkat cell adherence to non-activated TrkAIII-SH-SY5Y cells (upper left micrograph) not altered by Jurkat pre-incubation with BIRT377 (15 μM, upper right micrograph) compared to increased Jurkat cell adhesion to PMA-activated TrkAIII-SH-SY5Y cells (lower left micrographs) significantly reduced by Jurkat pre-incubation with BIRT377 (15 μM, lower right micrograph, bar = 10 μm). ( c ) Histogram demonstrating fold increase in Jurkat cell adhesion to PMA-activated TrkAIII-SH-SY5Y cells (PMA-act, Black column), with respect to Jurkat cell adhesion to non-activated TrkAIII-SH-SY5Y cells (Non-act, white column, arbitrary value of 1), and the significant reduction in the adhesion of Jurkat cells pre-incubated with BIRT377 (15 μM) to PMA-activated (PMA-act, column with white spots and black background) but not to non-activated TrkAIII-SH-SY5Y cells (Non-act, column with black spots and white background), in triplicate experiments, each performed in duplicate (* p < 0.0001).

Article Snippet: Rabbit polyclonal anti-human ICAM-1(CSB-PA07149A0Rb) and rabbit polyclonal anti-human CD11a (ITGAL CSB-PA011875LA01HU) antibodies were from Cusabio (Houston, TX, USA).

Techniques: Western Blot, Expressing, Staining, Incubation

Journal: International Journal of Molecular Sciences

Article Title: Jurkat T-Cell Antigen-Independent Elimination of PMA-Activated Neuroblastoma Cells Is Triggered by CCL2/CCR2, Depends Upon Lipid Raft LFA1/ICAM1 Immune Synapses, Is Mediated by m-TRAIL and Is Augmented by the TrkAIII Oncoprotein

doi: 10.3390/ijms27041970

Figure Lengend Snippet: Representative IF micrographs demonstrating clustering of Jurkat CD11a (arrows, red), TrkAIII-SH-SY5Y ICAM-1 (arrows, red) and DR5/TRAIL-R2 (arrows, green), relocalization of TrkAIII-SH-SY5Y Golgi (GM130, red/yellow) and centrosomes (γ-tubulin, green/yellow), and Jurkat TRAIL vesicle polarization (green) at interaction sites between Jurkat cells (J) and PMA-activated TrkAIII-SH-SY5Y cells (S) (Jurkat + PMA-act TrkAIII-SH-SY5Y) but not between Jurkat cells and non-activated TrkAIII-SH-SY5Y cells (Jurkat + non-act TrkAIII-SH-SY5Y) (nuclei are stained blue with DAPI; bar = 10 μm).

Article Snippet: Rabbit polyclonal anti-human ICAM-1(CSB-PA07149A0Rb) and rabbit polyclonal anti-human CD11a (ITGAL CSB-PA011875LA01HU) antibodies were from Cusabio (Houston, TX, USA).

Techniques: Staining