Journal: Oncology Letters

Article Title: Inhibitor of differentiation 1 in U87MG glioblastoma cells promotes HUVEC sprouting through endothelin-1

doi: 10.3892/ol.2022.13533

Figure Lengend Snippet: EDN1-EDNRB axis promotes endothelial sprouting, and its pro-angiogenesis function is more potent than that of VEGF. (A) HUVEC spheroids were cultured in the presence or absence of VEGF (50 ng/ml) and EDN1 (50 ng/ml) in EBM basal medium (red, phalloidin; blue, DAPI). Quantification of the sprouting length in HUVEC spheroids. ***P<0.001. Data are presented as the mean ± SEM. Scale bar, 100 µm. (B) HUVEC spheroids were cultured in the presence or absence of EDN1 (100 ng/ml) and bosentan (10 µM) (green, CD31). Quantification of the sprouting length in HUVEC spheroids. ***P<0.001. Data are presented as the mean ± SEM. Scale bar, 100 µm. (C) HUVEC spheroids were cultured in the presence or absence of EDN1 (100 ng/ml; green, EDNRA and EDNRB; red, phalloidin; blue, DAPI). Quantification of the fluorescence intensity (EDNRA and EDNRB) in the sprouting region. ***P<0.001. Data are presented as the mean ± SEM. Scale bar, 100 µm. EDN1, endothelin1; EDNRA, endothelin receptor type a; EDNRB, endothelin receptor type b; VEGF, vascular endothelial growth factor.

Article Snippet: The spheroids were incubated with CD31 antibody (1:200, Thermo Fisher, Cat. IHC-00055), EDNRA antibody (1:200, Alomone Labs, Cat. AER-001), and EDNRB antibody (1:200, Alomone Labs, Cat. AER-002) overnight at 4°C, followed by incubation with Alexa Fluor 488- or 568-conjugated secondary antibodies (1:400, ThermoFisher, Cat. A10042, A21202) for 2 h at RT.

Techniques: Cell Culture, Fluorescence

Journal: Neoplasia (New York, N.Y.)

Article Title: Irreversible and sustained upregulation of endothelin axis during oncogene-associated pancreatic inflammation and cancer

doi: 10.1016/j.neo.2019.11.001

Figure Lengend Snippet: Alterations in the endothelin receptors during cerulein-induced ADM in KC and WT mice. Triple color immunofluorescence was performed for studying the expression of ET A R (A) and ET B R (B) in the acinar and ductal cells. The ET-receptors are stained with purple fluorophore. Amylase (red) was used as a marker of acinar cells, while CK19 (green) was used as a marker for ductal cells. Representative images of the stained sections of the pancreas of WT and KC mice at 0, 2, 7, 21 days post-treatment with cerulein. (Scale bar = 20 µm;).

Article Snippet: The tissues were blocked with 2.5% horse serum and incubated with primary antibodies overnight diluted in PBS: anti-ET A R, 1:300, Alomone Labs, AER-001; anti-ET A R, 1:300, Santacruz, sc-135902; anti-ET B R, 1:300, Alomone labs, AER-002; anti-ET B R, 1:300, Santacruz, sc-21196; anti-CK19, 1:200, TROMA III, anti-F4/80, 1:80, e-Biosciences, 14-4801-82; anti-α-SMA, 1:200, Abcam, ab7817; anti-amylase 1:300, Sigma.

Techniques: Immunofluorescence, Expressing, Staining, Marker

Journal: Neoplasia (New York, N.Y.)

Article Title: Irreversible and sustained upregulation of endothelin axis during oncogene-associated pancreatic inflammation and cancer

doi: 10.1016/j.neo.2019.11.001

Figure Lengend Snippet: Increased infiltration of F4/80 positive macrophages is associated with both ET A R and ET B R expression in cerulein induced acute inflammation: Dual color immunofluorescence images of the pancreatic sections stained for ET A R and ET B R (red) and macrophage marker F4/80 (green) following cerulein and saline treatment mice at days 0, 2, 7 and 21. The representative images for ET A R (A) and ET B R (B) are depicted in panels A and C, respectively, with areas showing co-localization of ET-receptors with macrophage markers highlighted in box and zoomed in inset. The degree of overlap between F4/80 positive macrophages and ET A R or ET B R was measured using ImageJ using Manders overlap coefficients (B and D respectively). (** p < 0.005, ns = not significant) (Scale bar = 20 µm; zoom scale bar = 10 µm).

Article Snippet: The tissues were blocked with 2.5% horse serum and incubated with primary antibodies overnight diluted in PBS: anti-ET A R, 1:300, Alomone Labs, AER-001; anti-ET A R, 1:300, Santacruz, sc-135902; anti-ET B R, 1:300, Alomone labs, AER-002; anti-ET B R, 1:300, Santacruz, sc-21196; anti-CK19, 1:200, TROMA III, anti-F4/80, 1:80, e-Biosciences, 14-4801-82; anti-α-SMA, 1:200, Abcam, ab7817; anti-amylase 1:300, Sigma.

Techniques: Expressing, Immunofluorescence, Staining, Marker

Journal: Neoplasia (New York, N.Y.)

Article Title: Irreversible and sustained upregulation of endothelin axis during oncogene-associated pancreatic inflammation and cancer

doi: 10.1016/j.neo.2019.11.001

Figure Lengend Snippet: Increased infiltration of F4/80 positive macrophages is associated with ET B R expression in smoking induced chronic inflammation: Three color immunofluorescence images of the ET A R (purple), ET B R (red) and F4/80 (green) expression in the pancreas of WT and KC mice that were either sham treated or exposed to cigarette smoke. The representative images are shown in Panel A where the areas of overlap are highlighted in a box and zoomed (inset). The degree of overlap of F4/80 staining with ET A R and ET B R was measured using ImageJ to calculate Manders overlap coefficient (B and C respectively). (* p < 0.05, ** p < 0.005, ns = not significant) (Scale bar = 20 µm; zoom scale bar = 10 µm).

Article Snippet: The tissues were blocked with 2.5% horse serum and incubated with primary antibodies overnight diluted in PBS: anti-ET A R, 1:300, Alomone Labs, AER-001; anti-ET A R, 1:300, Santacruz, sc-135902; anti-ET B R, 1:300, Alomone labs, AER-002; anti-ET B R, 1:300, Santacruz, sc-21196; anti-CK19, 1:200, TROMA III, anti-F4/80, 1:80, e-Biosciences, 14-4801-82; anti-α-SMA, 1:200, Abcam, ab7817; anti-amylase 1:300, Sigma.

Techniques: Expressing, Immunofluorescence, Staining