Fig 1: Silencing of CD82 promote trophoblast outgrowth and migration in villous explant cultures.(A) Villous explants from 5 to 8 weeks of gestation were maintained in culture for 5 days on matrigel under low (3% O2) oxygen tension. Villous explants of CD82 siRNA significantly increase budding and outgrowth of EVT from the distal end of the villous tips comparing with negative control. Serial pictures of villous explants were taken under the light microscope after 24, 48, and 72 h of culture in vitro. (B) Villi (a) transfected with FITC-tagged siRNA, showing the transfection efficiency (b). Villi (c) as a control transfected nothing, showing the villi background fluorescence (d). (C) Three experiments as in B were quantified by measuring the the migration distance of villous tip relative to CON siRNA transfected for 24 h. (t-test).
Fig 2: Expression of CD82 in human placental villi and cell lines.(A) Immunostaining of CD82 in normal human placental villi in the first trimester, maternal decidua, second trimester and third trimester. (a) CD82 was strongly expressed in trophoblast columns (TC) and moderate in CTB of human placental villi during the first trimester. (c) CD82 was highly expressed in decidual cells (DC). (e) CD82 was highly expressed in the maternal decidua, not detected in EVT and very faint in anchoring villous during the early second trimester. Boxed region is enlarged on the upper panel. (g) CD82 was absent in villous and very faint in EVT during the late second trimester. (i) CD82 was moderate expressed in cytotrophoblast cells invaded into the maternal decidua and highly expressed in syncytiotrophoblast in the third trimester. (b, f, h, j) Immunohistochemical staining with anti-cytokeratin7 (CK7) as a marker of CTB, TC in the first trimester placental vill, EVT in the maternal decidua; (d) Immunohistochemical staining with anti-vimentin as a maker of decidual cells. (k, l, m, n) negative controls (NEG) on sections in which normal IgG was used in place of primary antibody. CTB: cytotrophoblast; STB: syncytiotrophoblast; TC: trophoblast column; EVT: extravillous trophoblast; MD: maternal decidua, AV: anchoring villous W: weeks of pregnancy; Bar represents 100 µm. (B) Expression of CD82 in different trophoblast cell lines determined by semiquantitative RT-PCR and Western blotting, respectively. GAPDH was used as an internal control for RT-PCR or loading control for Western blotting. HTR8/SVneo: a human invasive extravillous trophoblast cell line derived from immortalized first trimester trophoblast; B6Tert: immortalized cytotrophoblast cell line; JEG-3: human choriocarcinoma cell lines. (C) Immunofluorescence of CD82 in HTR8/SVneo cell lines. Fluorescence signals specific to CD82 antibody were visualized as green, and the nuclei were shown by DAPI staining (blue).
Fig 3: The relative optical density of CD9, CD81, CD82, and CD151 in bovine oocytes. Representative images of Western-blot analysis for targeted tetraspanins in bovine fresh (lane 1) and vitrified oocytes (lane 2). The relative density of individual tetraspanins was calculated as the ratio of the optical density of tetraspanins and actin antibodies signal in the blot. Bars represent the mean ± SEM of the density ratio of CD9, CD81, CD82, and CD151 to actin of three replicates. No statistically significant changes in protein levels in fresh vs. vitrified oocytes were revealed
Fig 4: Localization of tetraspanins CD151, CD82, and CD63 in fresh immature cow oocytes. Arrows point to CD151 (green) localization in clusters along the plasma membrane (PM) and/or in the perivitelline space (PVS) (a), CD82 (green) detection in clusters near the PM and zona pellucida (ZP) (arrows) (b), CD63 (green) detection in clusters near the PM and ZP (arrows) (c). DNA (blue). Images marked by letters with an apostrophe represent the bright field images (BF) corresponding to the fluorescence image marked with the same letter. The scale bar represents 50 µm
Fig 5: Localization of tetraspanins CD151, CD82, CD63, CD9, and CD81 in fresh and vitrified mature cow oocytes. Arrows show CD151 (green) localized in clusters along the plasma membrane (PM) in fresh mature (a) and vitrified mature oocytes (b), CD82 (green) detection in clusters near the PM and zona pellucida (ZP) in fresh mature (c) and vitrified mature oocytes (d), CD63 (green) localized to the PM and ZP of fresh mature (e) and vitrified mature oocytes (f), CD9 localization, detected by a monoclonal antibody, mCD9 (green), in PM and ZP in fresh mature (g) and vitrified mature oocytes (h). Detection of CD9 using a polyclonal antibody, pCD9 (green) in the PM with extension to the perivitelline space (PVS) in fresh mature (i) and vitrified mature oocytes (j), CD81 localization (green) in clusters in the PM of fresh mature (k) and vitrified mature oocytes (l). DNA (blue). Images marked by letters with an apostrophe represent the bright-field images (BF) corresponding to their respective fluorescence image. The scale bar represents 50 µm
Supplier Page from Abcam for Anti-CD82 antibody