Journal: Journal of Immunology Research

Article Title: Connexin 43 Communication Channels in Follicular Dendritic Cell Development and in Follicular Lymphomas

doi: 10.1155/2015/528098

Figure Lengend Snippet: Connexin 43 expression in secondary lymphoid follicles of reactive human tonsils. Cx43 immunoreaction (red) is accumulated mainly in the light zone localizing less Ki67 positive lymphocytes ((a), green) and more CD21 positive FDC processes ((b), green) than the dark zone (circled areas) of germinal center in consecutive sections. Cx43 (green) colocalizes with desmoplakin (red) produced by FDC ((c); arrowheads). Cx43 plaques (green) are also closely associated with B cells ((d); arrowheads) and rarely with CD4 positive T cells ((e); Cx43: red, CD4: green, arrowhead) in the germinal center. Immunofluorescence, nuclear staining in (a), (b), and (e) with Hoescht (blue) and in (d) with 7-aminoactinomycin D (red). LZ: light zone and MZ: mantle zone. Scale bar on (a) shows 30 µ m on (a) and (b); 15 µ m on (c) and (e); and 7 µ m on (d).

Article Snippet: After fixation in methanol-acetone (1 : 1) for 10 min, FDC-B cell cultures grown on microscope coverslips were also tested with immunofluorescence using rabbit anti-Cx43 (code: #3512, 1 : 200; Cell Signaling, Beverly, MA) and mouse anti-Ki67 (clone: Mib1, 1 : 100) IgG or combining either monoclonal mouse anti-Cx43 (Clone: CX-1B1, 1 : 100, Life Tech., Grand Island, NY) with rabbit anti-human IgM (A0425, 1 : 300), or IgG (A0423, 1 : 500) antibodies or rabbit anti-Cx43 (code: #3512, 1 : 200) with mouse anti-CD35 (clone: Ber-Mac-DRC, 1 : 50) antibodies for double labeling.

Techniques: Expressing, Produced, Immunofluorescence, Staining

Journal: Journal of Immunology Research

Article Title: Connexin 43 Communication Channels in Follicular Dendritic Cell Development and in Follicular Lymphomas

doi: 10.1155/2015/528098

Figure Lengend Snippet: Connexin 43 expression in follicular lymphomas (FL); lymph node localization ((a)–(f)); and bone marrow involvement ((g)–(j)). Large and irregular follicles dominated by CD10 positive FL B cells (a) and fragmented CD21 positive FDC meshwork (b). Cx43 ((c), red) reaction colocalizes with that of CD21 ((d), green) in yellow (arrowheads). Insets show a binuclear FDC with highly upregulated Cx43 ((c), green),and colocalization of Cx43 (red) and CD10 (green) as yellow dots (d). Elevated tumor cell proliferation detected with Ki67 reaction (green) is seen in FL follicles either with dense FDC and Cx43 reactions ((e); red) or with fragmented FDC and Cx43 staining ((f); circled area highlights broken or missing FDC). In the bone marrow involvement of FL NGFR (g) and CD21 (inset) positive follicles (arrowhead: LNGFR positive hyperplastic stromal cells) show the most of Cx43 reaction ((h), red) and proliferating tumor cells (Ki67-green). Strong Cx43 staining ((i), red) is accompanied with only fragmented CD21 reaction (green) in another follicle. Significantly more Cx43 (red) but less Ki67 (green) is detected within a bone marrow FL infiltrate than in the rest of bone marrow (j). Arrowheads in the inset show Cx43 within the boxed area. Graphs reveal linear correlation between Cx43 and CD21 levels in FL (k) and Cx43 (l) or Ki67 (m) expression in FDC rich and FDC poor FL areas. Immunoperoxidase ((a), (b), and (g)) and immunofluorescence staining ((c)–(f) and (h)–(j)). Blue nuclear staining using hematoxylin ((a), (b), and (g)) or Hoescht ((c)–(f) and (h)–(j)). Significance: * P < 0.05 and ** P < 0.005. Scale bar on (a) shows 150 µ m on (a) and (b); 30 µ m on (c) and (d); 120 µ m on (e), (f), (g), (i), and (j); and 100 µ m on (h).

Article Snippet: After fixation in methanol-acetone (1 : 1) for 10 min, FDC-B cell cultures grown on microscope coverslips were also tested with immunofluorescence using rabbit anti-Cx43 (code: #3512, 1 : 200; Cell Signaling, Beverly, MA) and mouse anti-Ki67 (clone: Mib1, 1 : 100) IgG or combining either monoclonal mouse anti-Cx43 (Clone: CX-1B1, 1 : 100, Life Tech., Grand Island, NY) with rabbit anti-human IgM (A0425, 1 : 300), or IgG (A0423, 1 : 500) antibodies or rabbit anti-Cx43 (code: #3512, 1 : 200) with mouse anti-CD35 (clone: Ber-Mac-DRC, 1 : 50) antibodies for double labeling.

Techniques: Expressing, Staining, Immunofluorescence

Journal: Journal of Immunology Research

Article Title: Connexin 43 Communication Channels in Follicular Dendritic Cell Development and in Follicular Lymphomas

doi: 10.1155/2015/528098

Figure Lengend Snippet: Testing of cell proliferation (gray columns) and proliferating cell fractions (numbers in gray columns in %) using Ki67 immunocytochemistry and absolute cell numbers (gray + white columns) indicating cell survival, in FDC-B cell cultures. Absolute cell numbers are significantly decreased ( * P < 0.05) 6 h, 10 h, and 16 h after Gap27 treatment compared to the untreated cultures (Unt). Proliferating cell fractions show a nonsignificant trend of reduction at these time points ( p 6 h = 0.423; p 10 h = 0.186; p 16 h = 0.067) in the control cultures. Results in graphs show the mean and standard deviation of at least three independent experiments.

Article Snippet: After fixation in methanol-acetone (1 : 1) for 10 min, FDC-B cell cultures grown on microscope coverslips were also tested with immunofluorescence using rabbit anti-Cx43 (code: #3512, 1 : 200; Cell Signaling, Beverly, MA) and mouse anti-Ki67 (clone: Mib1, 1 : 100) IgG or combining either monoclonal mouse anti-Cx43 (Clone: CX-1B1, 1 : 100, Life Tech., Grand Island, NY) with rabbit anti-human IgM (A0425, 1 : 300), or IgG (A0423, 1 : 500) antibodies or rabbit anti-Cx43 (code: #3512, 1 : 200) with mouse anti-CD35 (clone: Ber-Mac-DRC, 1 : 50) antibodies for double labeling.

Techniques: Immunocytochemistry, Standard Deviation