Journal: bioRxiv

Article Title: M. leprae infects human keratinocytes via the interaction of laminin-5 with α-dystroglycan, integrin-β1, or -β4

doi: 10.1101/591313

Figure Lengend Snippet: Human skin was immunostained with an antibody against LN-α2 or LN-5. After washing with PBS, the skin samples were incubated with a mouse Cy™5- or a rabbit Cy™5-conjugated secondary antibody at room temperature for 2 h. Nuclei were counterstained for 5 min with DAPI. Scale bar: 20 μm.

Article Snippet: Antibodies against LN-5 (ab102539 for immunohistochemistry), integrin-β1 (ab24693 for immunocytochemistry and binding assay) and -β4 (ab133682 for immunocytochemistry and binding assay) were obtained from Abcam (Cambridge, MA).

Techniques: Incubation

Journal: bioRxiv

Article Title: M. leprae infects human keratinocytes via the interaction of laminin-5 with α-dystroglycan, integrin-β1, or -β4

doi: 10.1101/591313

Figure Lengend Snippet: (A and B) M. leprae (5 × 10 8 ) were overlaid onto LN-α2, LN-5, collagen IV, or fibronectin-coated 4-chamber slides and incubated for 1 h at 37 °C. After removing unattached M. leprae by washing, the M. leprae were stained with Auramine O. The level of binding activity of M. leprae to ECM-coated slides was determined by measuring Auramine O fluorescence activity. * P <0.05 between the indicated groups. Scale bar: 20 μm.

Article Snippet: Antibodies against LN-5 (ab102539 for immunohistochemistry), integrin-β1 (ab24693 for immunocytochemistry and binding assay) and -β4 (ab133682 for immunocytochemistry and binding assay) were obtained from Abcam (Cambridge, MA).

Techniques: Incubation, Staining, Binding Assay, Activity Assay, Fluorescence

Journal: bioRxiv

Article Title: M. leprae infects human keratinocytes via the interaction of laminin-5 with α-dystroglycan, integrin-β1, or -β4

doi: 10.1101/591313

Figure Lengend Snippet: (A and B) HEKn cells were incubated with LN-α2- (A) or LN-5- (B) coated M. leprae at MOI of 10:1, 20:1, 50:1 and 100:1, respectively, for 1 h at 37 °C. After removing unattached M. leprae by washing, the samples were stained with the AFB stain. The number of M. leprae –bound cells per 100 HEKn cells was determined in the oil immersion field of light microscopy. * P <0.05 between the indicated groups.

Article Snippet: Antibodies against LN-5 (ab102539 for immunohistochemistry), integrin-β1 (ab24693 for immunocytochemistry and binding assay) and -β4 (ab133682 for immunocytochemistry and binding assay) were obtained from Abcam (Cambridge, MA).

Techniques: Incubation, Staining, Light Microscopy

Journal: bioRxiv

Article Title: M. leprae infects human keratinocytes via the interaction of laminin-5 with α-dystroglycan, integrin-β1, or -β4

doi: 10.1101/591313

Figure Lengend Snippet: (A) HEKn cells were pre-treated with an antibody against α-DG at the indicated dilution ratio for 1 h, and then further incubated with LN-α2-coated M. leprae at MOI of 100:1 for 1 h at 37 °C. (B and C) HEKn cells were pre-treated with an antibody against α-DG, integrin-β1, -β2, -β3, or -β4, at the dilution ratio 1:1,000 (B) and at the indicated dilution ratio (C) for 1 h. HEKn cells were then incubated with LN-5-coated M. leprae at MOI of 100:1 for 1 h at 37 °C. After unattached M. leprae were washed away, M. leprae were stained with the AFB stain. The number of M. leprae –bound cells per 100 HEKn cells was determined in the oil immersion field of light microscopy. * P <0.05 between the indicated groups.

Article Snippet: Antibodies against LN-5 (ab102539 for immunohistochemistry), integrin-β1 (ab24693 for immunocytochemistry and binding assay) and -β4 (ab133682 for immunocytochemistry and binding assay) were obtained from Abcam (Cambridge, MA).

Techniques: Incubation, Staining, Light Microscopy

Journal:

Article Title: Purification and Characterization of S -Adenosyl- l -Methionine: Desoxyhemigossypol-6- O- Methyltransferase from Cotton Plants. An Enzyme Capable of Methylating the Defense Terpenoids of Cotton 1

doi:

Figure Lengend Snippet: SDS-PAGE of the dHG-6-OMT active fraction from 2′,5′-ADP-Sepharose 4B affinity column (lane 1). Low-range silver-stain SDS-PAGE standards (Bio-Rad) (lane 2) are also shown.

Article Snippet: Ready gels (10%), the Silver-Stain Plus kit, and silver stain SDS-PAGE standard (low range) were purchased from Bio-Rad (Hercules, CA).

Techniques: SDS Page, Affinity Column, Silver Staining