chicken igy isotype  (R&D Systems)

Journal: eLife

Article Title: TLR2 regulates hair follicle cycle and regeneration via BMP signaling

doi: 10.7554/eLife.89335

Figure Lengend Snippet: The promotion of hair follicle regeneration after wound healing is dependent on TLR2. ( A ) Representative confocal images of Nile red-labeled (sebaceous gland) wild-type (WT) telogen hair follicles co-immunostained for MPO showing complete co-localization of MPO to the sebaceous gland. The isotype control panel shows the images of hair follicles stained with MPO isotype control antibody. Scale bars are 20 μm. ( B ) Representative confocal images of hair follicles from old vs young mice stained for MPO. Scale bars are 10 μm. ( C ) Quantification of MPO fluorescent intensity in B showing significantly less MPO in hair follicles from older mice. N=6 for each group. ( D ) Representative microphotographs of human hair follicle stem cells (HFSCs) pre-treated with 10 µg/ml MAb-mTLR2 or DMSO and co-cultured with/without 2.5 µM of CEP. Representative images from at least three independent assays are shown. Scale bar 50 µm. ( E ) Bar graphs show increased proliferation of HFSC in the presence of TLR2 endogenous ligand CEP compared to control, which was abolished in the presence of TLR2 blocking antibody. N=6 independent experiments. ( F ) Bar graphs show increased proliferation of human hair follicle dermal papilla cells incubated with 5 µM of CEP compared to the control. N=9 independent experiments. ( G ) Representative confocal images of Ki67 immunostaining of dorsal skin adjacent to wound of CEP-treated WT bone marrow transplanted WT and TLR2 KO mice. Scale bars are 50 μm. ( H ) Quantitative results showed increased Ki67 intensity in hair follicles around wounds of CEP-treated WT bone marrow transplanted WT mice with no differences in TLR2 KO with WT bone marrow. N=4 per group. ( I ) Representative photographs of dorsal skin (upper panels), inner skin flaps (middle panels), and representative confocal images of Ki67 immunostaining (lower panels) of vehicle- or CEP-treated WT or TLR2 KO skin. Scale bars are 1 mm for dorsal skin, 500 μm for skin flaps, and 50 μm for confocal images. ( J ) Bar graph showing quantification of hair follicle numbers of vehicle or CEP-treated skin from I. N=5 per group. ( K ) Bar graph showing quantification of Ki67 fluorescent intensity of Ki67 staining of vehicle or CEP-treated skin from I. N=4 per group. Unpaired two-tailed t-test ( C ), or non-parametric Mann-Whitney test ( H, J, K ), or Kruskal-Wallis test with Dunn’s multiple comparisons test ( F ), or one-way ANOVA with Tukey’s multiple comparisons test ( E ) was used to determine statistical differences. All bar graphs are mean ± s.e.m. A p-value ≤ 0.05 was considered to be statistically significant.

Article Snippet: Antibody , Chicken IgY Isotype Control , Novus Biologicals , Cat.# AB-101-C RRID: AB_354263 , According to immune antibody concentration.

Techniques: Labeling, Control, Staining, Cell Culture, Blocking Assay, Incubation, Immunostaining, Two Tailed Test, MANN-WHITNEY

Journal: eLife

Article Title: TLR2 regulates hair follicle cycle and regeneration via BMP signaling

doi: 10.7554/eLife.89335

Figure Lengend Snippet:

Article Snippet: Antibody , Chicken IgY Isotype Control , Novus Biologicals , Cat.# AB-101-C RRID: AB_354263 , According to immune antibody concentration.

Techniques: Activity Assay, Knock-In, Sequencing, Isolation, Blocking Assay, Control, Concentration Assay, Staining, Membrane, Recombinant, Software

Journal: Frontiers in Cell and Developmental Biology

Article Title: Possible Therapeutic Utility of anti-Cell Adhesion Molecule 1 Antibodies for Malignant Pleural Mesothelioma

doi: 10.3389/fcell.2022.945007

Figure Lengend Snippet: Immunofluorescence of CADM1 in MESO-14 and NCI-H2052 cells cultured on a MeT-5A cell monolayer in the presence of 9D2 and 3E1. MESO-14 and NCI-H2052 cell sheets were labeled with DiI (red) and were cocultured on a MeT-5A cell monolayer in the presence of control IgY (1.0 μg/ml; left), 9D2 (1.0 μg/ml; middle), or 9D2 and 3E1 (1.0 μg/ml each; right). After 2 days, the cocultures were immunostained with the anti-CADM1 antibody (green). Nuclei were counterstained with DAPI (blue). Images were captured by a confocal microscope; representatives are shown. Bar = 50 μm. In each photomicrograph, a boxed area is enlarged at the bottom for observation at single-cell level.

Article Snippet: A control chicken IgY was purchased from R&D Systems (Minneapolis, MN, United States), and a normal human IgG from Fujifilm Wako Chemicals (Osaka, Japan).

Techniques: Immunofluorescence, Cell Culture, Labeling, Microscopy

Journal: Frontiers in Cell and Developmental Biology

Article Title: Possible Therapeutic Utility of anti-Cell Adhesion Molecule 1 Antibodies for Malignant Pleural Mesothelioma

doi: 10.3389/fcell.2022.945007

Figure Lengend Snippet: Growth suppression of MPM cells on MeT-5A cells by 9D2 and 3E1 (A) MESO-14 and NCI-H2052 cell sheets were labeled with DiI (red) and were cocultured on a MeT-5A cell monolayer in the presence of control IgY (1.0 μg/ml; left), 9D2 (1.0 μg/ml; middle), or 9D2 and 3E1 (1.0 μg/ml each; right). Cell sheets were observed through a fluorescence microscope at the time of antibody addition (0 d) and after 2 days (2 d) (B) MESO-1, MESO-1-CADM1, NCI-H28, NCI-H2052, and MESO-14 cell lines were cocultured on a MeT-5A cell monolayer in the presence of either control IgY (1.0 μg/ml) or 9D2 (1.0 μg/ml) (upper graph), and in the presence of either control IgY (2.0 μg/ml) or 9D2 and 3E1 (1.0 μg/ml each) (lower graph). For individual cell sheets, the ratio of the cell number at the time of antibody addition (0 d) to that after 2 days (2 d) was calculated, and plotted as the growth rate in a dot graph. The mean was calculated from the ratios of five to ten sheets for each coculture group, and is shown below or above the dots. p -values are shown at the top of each graph. Bar = 100 μm (C) TUNEL assays of MESO-14 and NCI-H2052 cells cocultured on a MeT-5A cell monolayer for 2 days in the presence of either control IgY or 9D2 and 3E1. The mean proportion of TUNEL-positive MPM cells (%) is plotted with a thin line indicating the standard deviation. A p -value between two groups is shown at the top of each graph.

Article Snippet: A control chicken IgY was purchased from R&D Systems (Minneapolis, MN, United States), and a normal human IgG from Fujifilm Wako Chemicals (Osaka, Japan).

Techniques: Labeling, Fluorescence, Microscopy, TUNEL Assay, Standard Deviation