Journal: Journal of visualized experiments : JoVE

Article Title: Retroviral overexpression of CXCR4 on murine B-1a cells and adoptive transfer for targeted B-1a cell migration to the bone marrow and IgM production

doi: 10.3791/61003

Figure Lengend Snippet: Table of Materials

Article Snippet: Incubate on ice for 10 minutes at 4 °C. table ft1 table-wrap mode="anchored" t5 caption a7 Name of Material/Equipment Company Catalogue Number Comments/Description anti-CD16/CD32, or Fc block Life Technologies MFCR00 Ter119 biotin eBioscience 13–5921–82 Clone: Ter119 CD3e biotin eBioscience 13–0033–85 Clone: eBio500A2 Gr-1 biotin eBioscience 13–5931–82 Clone: RB6–8C5 CD23 biotin eBioscience 13–0232–81 Clone: B3B4 NK1.1 biotin BD Biosciences 553163 Clone: PK136 F4/80 biotin Life Technologies MF48015 Clone: BM8 anti-biotin microbeads Miltenyi Biotec 130–090–485 LS columns Miltenyi Biotec 130–042–401 CD5 PE eBioscience 12–0051–83 Clone: 53–7.3 IgM PECF594 BD Biosciences 562565 Clone: R6–60.2 CD23 PECy7 eBioscience 25–0232–82 Clone: B3B4 B220 APC eBioscience 17–0452–83 Clone: RA3–6B2 CD19 APCef780 eBioscience 47–0193–82 Clone: eBio1D3 CD45.1 PerCP-Cy5.5 BD Biosciences 560580 Clone: A20 CD45.2 BV421 BD Biosciences 562895 Clone: 104 CXCR4 APC eBioscience 17–9991–82 Clone: 2B11 Live/Dead Yellow Life Technologies {"type":"entrez-nucleotide","attrs":{"text":"L34968","term_id":"522211","term_text":"L34968"}} L34968 RPMI-1640 Gibco 11875–093 heat-inactivated fetal bovine serum Gibco 16000–044 HEPES Gibco 15630–080 Non-essential amino acids Gibco 11140–050 Sodium pyruvate Gibco 11360–070 Gentamicin Gibco 15710–064 Beta-mercaptoethanol Gibco 21985–023 ODN 1668 InvivoGen tlrl-1668 70 micron filter caps Falcon 352235 Insulin syringes BD Biosciences 329461 Ctl-GFP retrovirus N/A N/A Generated in house using GFP-expressing retroviral plasmid MigR1 provided by Dr. T.P.

Techniques: Blocking Assay, Generated, Retroviral, Plasmid Preparation, Cloning, Software

Journal: bioRxiv

Article Title: Loss of cyclin A2 in murine colonic epithelial cells disrupts colon homeostasis by triggering DNA damage and dysplasia and high cyclin A2 expression is a good-prognosis factor in patients with colorectal cancer

doi: 10.1101/690404

Figure Lengend Snippet: (A). Cyclin A2 depletion at the protein level in colonic epithelial cells (CEC) of constitutive ( VilCreCcna2fl/fl , left panel) and tamoxifen-induced knockout mice ( VilCreERT2Ccna2fl/fl , right panel) by comparison to control animals ( Ccna2fl/fl ) displayed by Western Blot analysis. ( B, C, D). Immunostaining for cyclin A2 ( B ), F4/80 ( C ) and IL-6 ( D ) of the indicated parts of colon derived from control ( Ccna2fl/fl ), and constitutive ( VilCreCcna2fl/fl ) cyclinA2-deficient mice. Blow Up: 2.5X. Scale bars: 100μm.

Article Snippet: The following antibodies were used: rabbit anti-Cyclin A2 (Abcam, ab181591, 1 :1000 dilution), rabbit anti-Ki67 (Abcam, ab16667, 1 :200 dilution), rat anti-F4/80 (Hycult biotech, HM1066 1 :800 dilution), rabbit anti-non-phospho (Active) β-catenin (Cell signaling, #8814, 1 :1000 dilution), rabbit anti-γH2AX (Abcam, ab11174, 1 :500), rat anti-IL6 (Novus Biological, IL6/1270, 1:100), rabbit anti-TAZ (Novus Biological, NBP1-85067, 1:100).

Techniques: Knock-Out, Western Blot, Immunostaining, Derivative Assay