Journal:

Article Title: Interaction of Mouse Polycomb-Group (Pc-G) Proteins Enx1 and Enx2 with Eed: Indication for Separate Pc-G Complexes

doi:

Figure Lengend Snippet: Immunolocalization of HA-Eed, HA-ENX1, and the Pc-G complex protein Mph1 in U2-OS cells. (Top) U2-OS cells transfected with the (HA)3-ENX1 construct were fixed and incubated with both the 12CA5 monoclonal antibody and an affinity-purified rabbit anti-Mph1 serum. Detection was carried out with FITC-conjugated anti-mouse IgG and TRITC-conjugated anti-rabbit IgG, using confocal laser-scanning microscopy. The FITC signal (left) indicates a diffuse nuclear staining together with concentrations in subnuclear domains for ENX1. The TRITC signal (middle) detects endogenous expression of human MPH1 in subnuclear domains, characteristic of a previously described human Pc-G multiprotein complex (3). The merged image (right) shows a partial colocalization of ENX1 and MPH1 domains (ENX1 in green, MPH1 in red, overlap in yellow/orange). (Bottom) U2-OS cells transfected with the HA-Eed expression construct were fixed and stained with 12CA5 and anti-Mph1 sera as above. Eed is localized diffusely throughout the nucleus. Note the specificity of the 12CA5 serum for transfected cells: in two untransfected cells (merged images), only the TRITC signal is detected. Bar, 10 μm.

Article Snippet: After being washed, the membrane was incubated for 1 h at room temperature with horseradish peroxidase-linked goat anti-rabbit immunoglobulin G IgG (Biosource) or goat anti-mouse IgG (Bio-Rad) as the second antibody diluted 1:10,000 in PBST containing 1% dried milk.

Techniques: Transfection, Construct, Incubation, Affinity Purification, Confocal Laser Scanning Microscopy, Staining, Expressing

Journal: Molecular human reproduction

Article Title: Ectopic endometrial cells express high concentrations of interleukin (IL)-8 in vivo regardless of the menstrual cycle phase and respond to oestradiol by up-regulating IL-1-induced IL-8 expression in vitro.

doi: 10.1093/molehr/7.9.859

Figure Lengend Snippet: Figure 1. Localization of interleukin-8 (IL-8) in endometriotic lesions by immunohistochemistry. Sections were incubated with polyclonal rabbit anti-IL-8 antibody, then with biotin-conjugated anti-rabbit antibody and streptavidin–peroxidase. The immunoreaction was revealed with DAB as chromogen, and haematoxylin which stains the nucleus in blue was used for counterstaining. Note the intense brown positive immunostaining in endometriotic glands and stroma from (A) an ovarian endometriotic lesion, stage IV, proliferative phase, day 4 (patient no. 6, Table I), and (B) the inner lining of ovarian endometrioma, stage IV, secretory phase, day 22 (patient no. 10, Table I). (C and D) control sections incubated with normal rabbit IgG instead of the primary polyclonal rabbit antibody showing no immunostaining. Scale bar 50 µm.

Article Snippet: Immunostaining was performed using a polyclonal rabbit anti-IL-8 antibody [10 μg/ml in PBS containing 1% bovine serum albumin (BSA)] (Biosource International, Camarillo, CA, USA), a biotin-conjugated affinity-purified goat anti-rabbit antibody (H L) (2 μg/ml) (Jackson ImmunoResearch Laboratories Inc., West Grove, PA, USA), peroxidase-conjugated streptavidin (1.3 μg/ml) (Jackson ImmunoResearch Laboratories), diaminobenzidine (Sigma Chemical Co, St Louis, MO, USA) as chromogen and haematoxylin for counterstaining.

Techniques: Immunohistochemistry, Incubation, Immunostaining, Control