rabbit polyclonal anti cleaved caspase 9  (Cell Signaling Technology Inc)

Journal: BMC Medicine

Article Title: α-Mangostin extracted from the pericarp of the mangosteen ( Garcinia mangostana Linn) reduces tumor growth and lymph node metastasis in an immunocompetent xenograft model of metastatic mammary cancer carrying a p53 mutation

doi: 10.1186/1741-7015-9-69

Figure Lengend Snippet: Apoptosis and cell cycle in vitro study (mouse mammary carcinoma BJMC3879luc2 cells) . (A) Cell viability was significantly lower in mouse mammary carcinoma BJMC3879luc2 cells treated with more than 12 μM α-mangostin for 24 or 48 hours (** P < 0.01). Five samples from each dosage of α-mangostin were examined. The IC 50 concentration was determined to be 12 μM; therefore, 12 μM α-mangostin and 24 hour-incubation was used for all in vitro studies. ( B) Caspase activities were evaluated using the luminescence assay. Activities of caspase-3, caspase-8 and caspase-9, but not caspase-12, were significantly elevated in BJMC3879luc2 cells treated with 12 μM α-mangostin for 24 hours (* P < 0.05 or ** P < 0.01). Six samples from the control and eight samples from α-mangostin-treated cells were used for measurement of caspase-3 activity, and three samples from each group were used for activity measurements of the other caspases. ( C) Cytochrome c in the cytosolic fraction, as determined by ELISA, was significantly increased in BJMC3879luc2 cells treated with α-mangostin for 24 hours compared to control levels (* P < 0.05). Three samples each from the control and α-mangostin-treated cell cultures were examined. ( D) Western blots of Bid (22 kDa) in BJMC3879luc2 cells treated with or without α-mangostin for 24 hours were similar (upper panel). Cleaved Bid (15 kDa) was not observed after α-mangostin treatment. β-Actin served as the internal control (lower panel). ( E) In BJMC3879luc2 cells treated with α-mangostin for 24 hours, cell viability was significantly increased by 10 or 100 μM of the broad-spectrum caspase inhibitor z-VAD-fmk, the caspase-3 specific inhibitor Ac-DNLD-CHO, the caspase-8 specific inhibitor z-IETD-fmk, and the caspase-9 specific inhibitor z-LETD-fmk (** P < 0.01). ( F) Cell-cycle analysis showed that α-mangostin induced arrest in the G1-phase and inhibition of cells entering the S-phase in metastatic mouse mammary carcinoma BJMC3879luc2 cells (** P < 0.01). Data are presented as mean ± SD. Ac: acetyl; CHO: aldehyde; ELISA: enzyme-linked immunosorbent assay; fmk: fluoromethyl ketone; z: N-benzyloxycarbonyl.

Article Snippet: Active caspase expression in the mammary tumor tissues was immunohistochemically detected using anti-cleaved caspase-3 and anti-cleaved caspase-9 rabbit polyclonal antibodies (Cell Signaling Technology).

Techniques: In Vitro, Concentration Assay, Incubation, Luminescence Assay, Activity Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Cell Cycle Assay, Inhibition

Journal: BMC Medicine

Article Title: α-Mangostin extracted from the pericarp of the mangosteen ( Garcinia mangostana Linn) reduces tumor growth and lymph node metastasis in an immunocompetent xenograft model of metastatic mammary cancer carrying a p53 mutation

doi: 10.1186/1741-7015-9-69

Figure Lengend Snippet: Apoptosis, active-caspase expression, angiogenesis and dilated lymphatic vessels with cancer cell invasion in mammary carcinomas . TUNEL-positive apoptotic cells are much more frequently seen in the tumor of a mouse given 20 mg/kg/day α-mangostin (B) than in the tumor of a control mouse (A) . Expression of active caspase-3 (C, D) and active caspase-9 (E, F) was more prominent in the tumor of a mouse given 20 mg/kg/day α-mangostin (D, F) than in a control mouse (C, E) . The number of CD-31-positive endothelial cells was lower in the tumor of a mouse given 20 mg/kg/day α-mangostin (H) compared to that of a control mouse (G) . Podoplanin-positive lymphatic vessels of a tumor in a control mouse were often dilated and filled with tumor cells ( arrow, I) . Mammary cancer cells were also observed in the intraluminal space of the dilated lymphatic vessels in the tumor of a mouse given 20 mg/kg/day α-mangostin ( arrow, J) . (A-H) : Scale bar = 50 μm; (I, J) : Scale bar = 25 μm. (A, B) : TUNEL stain; (C, D) : active caspase-3 immunohistochemistry ; (E, F) : active caspase-9 immunohistochemistry ; (G, H) : CD31 immunohistochemistry ; (I, J) : podoplanin immunohistochemistry .

Article Snippet: Active caspase expression in the mammary tumor tissues was immunohistochemically detected using anti-cleaved caspase-3 and anti-cleaved caspase-9 rabbit polyclonal antibodies (Cell Signaling Technology).

Techniques: Expressing, TUNEL Assay, Staining, Immunohistochemistry