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  • 99
    Cell Signaling Technology Inc cleaved caspase 3
    REV-ERB agonist SR9011 inhibit autophagy a–b, SR90011 treatment reduces the number of autophagosomes both in MCF7 and T47D, n= biological independent samples MCF7 20µM 24h n=9 (mock), n=4 (SR9011) ** P =0.0056, T47D 48h 20µM n=5 (mock), n=4 (SR9011) ** P =0.0079; c–d, SR9011 induces accumulation of p62 as shown by immunofluorescence both in MCF7 and T47D n= biological independent samples 48h MCF7 p62 n=3 (mock), n=4 (SR9011) * P =0.0286; 48h T47D n=5 (mock, SR9011) ** P =0.004; e, Accumulation of p62 is confirmed by immunoblot (48h, 20µM A375); f–g, Inhibition of autophagy precedes apoptosis induction as shown by immunofluorescence of p62, cleaved <t>Caspase</t> 3 and TUNEL assay (n= biological independent samples n=4 mock 48h, n=5 SR9011 p62, n=3 48h SR9011; n=6 (mock, SR9011 72h p62), n=10 (mock 72h), n=8 (SR9011 72h) A375 20µM, Cl. Casp. 3 48h * P =0.0286; Cl. Casp. 3 72h **** P
    Cleaved Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 38170 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti cleaved caspase 3
    Knockout of claudin-1 inhibits proliferation of cervical adenocarcinoma cells. (A-D) CLDN1 KO significantly inhibited proliferation of OMC4 cells in multiple assays, including cell count (A), colony formation assay (B), and immunostaining with anti–Ki-67 (proliferation marker) antibody (C). (D) Immunostaining with anti–cleaved caspase 3 (apoptosis marker) antibody. * P
    Anti Cleaved Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 11375 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc caspase 3
    Evaluation of V-9302  in vivo  in HCC1806 cell line xenograft-bearing mice ( A ) Tumor volumetric analysis of mice treated with vehicle or V-9302 (75 mg/kg per day) for 10 days.  n  = 5 mice per group. Treatment started 9 days post injection. P value at day 10 determined by Student’s  t  test. ( B ) Immunofluorescence analysis of tumor tissues harvested from vehicle- or V-9302-treated mice; LC3B and pAKT (Ser473) shown (pink). CD-31 positive vessels shown in green and nuclei in blue (DAPI). 20× magnification. P values determined by Student’s  t  test. ( C ) Effects of V-9302 treatment on pS6-positive cells and caspase 3-positive cells by immunohistochemistry. Quantitative analysis consisted of the mean counts of at least three representative fields from three vehicle and three V-9302-treated mice. For box plots, center line is plotted at the median; the box spans from the first quartile to the third quartile; whiskers represent min to max. Error bars represent ± std. dev.
    Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 28224 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc rabbit anti cleaved caspase 3
    AgNP-induced occurrence of systemic apoptosis. Immunofluorescent signals of the apoptotic biomarker “active caspase 3” in multiple tissues (brain, salivary gland, wing disc and gut) dissected from AgNP-exposed 3 rd -instar wandering larvae.
    Rabbit Anti Cleaved Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 5791 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti cleaved caspase 3 antibody
    p53 25,26,53,54/+  Embryos Exhibit Additional Features of CHARGE Syndrome and p53-Dependent Cellular Responses (a)  Double outlet right ventricle (DORV) in E13.5 p53 25,26,53,54/+  heart (50%, n=6). Top: Main pulmonary artery (MPA) connects via pulmonary valve (PV) to right ventricle (RV) in both control and p53 25,26,53,54/+  embryo. Bottom: Aorta (Ao) in control embryo connects to left ventricle (LV) via aortic valve (AV) Φ . Aorta in p53 25,26,53,54/+  embryo connects to RV via AV*. (b)  Abnormal atrioventricular cushions in E13.5 p53 25,26,53,54/+  heart (75%, n=4) fail to elongateinto mature mitral (mv, arrowhead) and tricuspid (tv, arrow) valves. RA: right atrium; LA: left atrium.  (c)  E13.5  p53 25,26,53,54/+  kidneys are smaller (79%), with fewer average glomeruli (13 vs. 3; n=5; arrows), than controls. (d) p53 25,26,53,54/+  embryonic phenotypes observed in CHARGE (+present, −absent).  (e)  Left: Cleaved-caspase 3 (CC3; Top) and p53 (Bottom) immunohistochemistry in E15.5 retinas. Arrows: CC3-positive cells. Right: CC3-positive cells per retinal area. ***p-value=0.007; one-tailed Welsh’s t-test (n=5).  (f)  BrdU immunofluorescence in E9.5 Pax3 +  NCCs (delineated by green-dotted line;  Extended-Data Fig. 6c ). Right: Percentage BrdU-positive cells per total Pax3 +  NCCs ***p-value=0.004 one-tailed Student’s t-test (n=4).
    Anti Cleaved Caspase 3 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1358 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti cleaved caspase3
    p53 25,26,53,54/+  Embryos Exhibit Additional Features of CHARGE Syndrome and p53-Dependent Cellular Responses (a)  Double outlet right ventricle (DORV) in E13.5 p53 25,26,53,54/+  heart (50%, n=6). Top: Main pulmonary artery (MPA) connects via pulmonary valve (PV) to right ventricle (RV) in both control and p53 25,26,53,54/+  embryo. Bottom: Aorta (Ao) in control embryo connects to left ventricle (LV) via aortic valve (AV) Φ . Aorta in p53 25,26,53,54/+  embryo connects to RV via AV*. (b)  Abnormal atrioventricular cushions in E13.5 p53 25,26,53,54/+  heart (75%, n=4) fail to elongateinto mature mitral (mv, arrowhead) and tricuspid (tv, arrow) valves. RA: right atrium; LA: left atrium.  (c)  E13.5  p53 25,26,53,54/+  kidneys are smaller (79%), with fewer average glomeruli (13 vs. 3; n=5; arrows), than controls. (d) p53 25,26,53,54/+  embryonic phenotypes observed in CHARGE (+present, −absent).  (e)  Left: Cleaved-caspase 3 (CC3; Top) and p53 (Bottom) immunohistochemistry in E15.5 retinas. Arrows: CC3-positive cells. Right: CC3-positive cells per retinal area. ***p-value=0.007; one-tailed Welsh’s t-test (n=5).  (f)  BrdU immunofluorescence in E9.5 Pax3 +  NCCs (delineated by green-dotted line;  Extended-Data Fig. 6c ). Right: Percentage BrdU-positive cells per total Pax3 +  NCCs ***p-value=0.004 one-tailed Student’s t-test (n=4).
    Anti Cleaved Caspase3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 595 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cleaved caspase3/product/Cell Signaling Technology Inc
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    Cell Signaling Technology Inc caspase 9
    p53 25,26,53,54/+  Embryos Exhibit Additional Features of CHARGE Syndrome and p53-Dependent Cellular Responses (a)  Double outlet right ventricle (DORV) in E13.5 p53 25,26,53,54/+  heart (50%, n=6). Top: Main pulmonary artery (MPA) connects via pulmonary valve (PV) to right ventricle (RV) in both control and p53 25,26,53,54/+  embryo. Bottom: Aorta (Ao) in control embryo connects to left ventricle (LV) via aortic valve (AV) Φ . Aorta in p53 25,26,53,54/+  embryo connects to RV via AV*. (b)  Abnormal atrioventricular cushions in E13.5 p53 25,26,53,54/+  heart (75%, n=4) fail to elongateinto mature mitral (mv, arrowhead) and tricuspid (tv, arrow) valves. RA: right atrium; LA: left atrium.  (c)  E13.5  p53 25,26,53,54/+  kidneys are smaller (79%), with fewer average glomeruli (13 vs. 3; n=5; arrows), than controls. (d) p53 25,26,53,54/+  embryonic phenotypes observed in CHARGE (+present, −absent).  (e)  Left: Cleaved-caspase 3 (CC3; Top) and p53 (Bottom) immunohistochemistry in E15.5 retinas. Arrows: CC3-positive cells. Right: CC3-positive cells per retinal area. ***p-value=0.007; one-tailed Welsh’s t-test (n=5).  (f)  BrdU immunofluorescence in E9.5 Pax3 +  NCCs (delineated by green-dotted line;  Extended-Data Fig. 6c ). Right: Percentage BrdU-positive cells per total Pax3 +  NCCs ***p-value=0.004 one-tailed Student’s t-test (n=4).
    Caspase 9, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 8293 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    REV-ERB agonist SR9011 inhibit autophagy a–b, SR90011 treatment reduces the number of autophagosomes both in MCF7 and T47D, n= biological independent samples MCF7 20µM 24h n=9 (mock), n=4 (SR9011) ** P =0.0056, T47D 48h 20µM n=5 (mock), n=4 (SR9011) ** P =0.0079; c–d, SR9011 induces accumulation of p62 as shown by immunofluorescence both in MCF7 and T47D n= biological independent samples 48h MCF7 p62 n=3 (mock), n=4 (SR9011) * P =0.0286; 48h T47D n=5 (mock, SR9011) ** P =0.004; e, Accumulation of p62 is confirmed by immunoblot (48h, 20µM A375); f–g, Inhibition of autophagy precedes apoptosis induction as shown by immunofluorescence of p62, cleaved Caspase 3 and TUNEL assay (n= biological independent samples n=4 mock 48h, n=5 SR9011 p62, n=3 48h SR9011; n=6 (mock, SR9011 72h p62), n=10 (mock 72h), n=8 (SR9011 72h) A375 20µM, Cl. Casp. 3 48h * P =0.0286; Cl. Casp. 3 72h **** P

    Journal: Nature

    Article Title: Pharmacological activation of REV-ERBs is lethal in cancer and oncogene induced senescence

    doi: 10.1038/nature25170

    Figure Lengend Snippet: REV-ERB agonist SR9011 inhibit autophagy a–b, SR90011 treatment reduces the number of autophagosomes both in MCF7 and T47D, n= biological independent samples MCF7 20µM 24h n=9 (mock), n=4 (SR9011) ** P =0.0056, T47D 48h 20µM n=5 (mock), n=4 (SR9011) ** P =0.0079; c–d, SR9011 induces accumulation of p62 as shown by immunofluorescence both in MCF7 and T47D n= biological independent samples 48h MCF7 p62 n=3 (mock), n=4 (SR9011) * P =0.0286; 48h T47D n=5 (mock, SR9011) ** P =0.004; e, Accumulation of p62 is confirmed by immunoblot (48h, 20µM A375); f–g, Inhibition of autophagy precedes apoptosis induction as shown by immunofluorescence of p62, cleaved Caspase 3 and TUNEL assay (n= biological independent samples n=4 mock 48h, n=5 SR9011 p62, n=3 48h SR9011; n=6 (mock, SR9011 72h p62), n=10 (mock 72h), n=8 (SR9011 72h) A375 20µM, Cl. Casp. 3 48h * P =0.0286; Cl. Casp. 3 72h **** P

    Article Snippet: Apoptosis was evaluated by immunostaining of cleaved caspase 3 (Cell signaling #9664 1:200) and by TUNEL assay using In Situ Cell Death Detection Kit, Fluoresceine or TMR red (Roche).

    Techniques: Immunofluorescence, Inhibition, TUNEL Assay

    SR9009 and SR9011 treatment evokes an apoptotic response and induces inhibition of autophagy in OIS cells a, Proliferation assay shows that REV-ERBs agonists impair viability of OIS cells (6 days, 20µM). b–c, Immunofluorescence assay for cleaved Caspase 3 and TUNEL assay shows apoptosis induction specifically in OIS (n=biological independent samples, n=7 mock, n=9 SR9009, n=14 SR9011, 72h, 20µM; one-way ANOVA, Cl. Casp 3 **** P

    Journal: Nature

    Article Title: Pharmacological activation of REV-ERBs is lethal in cancer and oncogene induced senescence

    doi: 10.1038/nature25170

    Figure Lengend Snippet: SR9009 and SR9011 treatment evokes an apoptotic response and induces inhibition of autophagy in OIS cells a, Proliferation assay shows that REV-ERBs agonists impair viability of OIS cells (6 days, 20µM). b–c, Immunofluorescence assay for cleaved Caspase 3 and TUNEL assay shows apoptosis induction specifically in OIS (n=biological independent samples, n=7 mock, n=9 SR9009, n=14 SR9011, 72h, 20µM; one-way ANOVA, Cl. Casp 3 **** P

    Article Snippet: Apoptosis was evaluated by immunostaining of cleaved caspase 3 (Cell signaling #9664 1:200) and by TUNEL assay using In Situ Cell Death Detection Kit, Fluoresceine or TMR red (Roche).

    Techniques: Inhibition, Proliferation Assay, Immunofluorescence, TUNEL Assay

    REV-ERBs agonist SR9009 inhibits autophagy a–b , SR9009 treatment reduces the number of autophagosomes, as shown by immunofluorescence of LC3B, (n=biological independent samples MCF7 (n=6 mock), (n=5 SR9009) and T47D (n=5 mock) (n=4 SR9009) Mann–Whitney test one-tailed MCF7 20µM 24h * P =0.0152, T47D 20µM 48h ** P =0.0079; c–d SR9009 induces accumulation of p62 as shown by immunofluorescence; n= biological independent samples MCF7 (n=3 mock), (n=8 SR9009) and T47D (n=5 mock), (n=4 SR9009) Mann–Whitney test one-tailed 48h MCF7 p62 ** P =0.0061; 48h T47D ** P =0.0079; e, Inhibition of autophagy is confirmed by the immunoblot for p62 (20µM 48h, A375); f–g, Inhibition of autophagy precedes apoptosis induction as shown by immunofluorescence of p62, cleaved Caspase 3 and TUNEL assay; n=biological independent samples, Mann–Whitney test one-tailed, A375 20µM Cl. Casp. 3 48h (n=3) * P =0.0179; Cl. Casp. 3 72h (n=7) **** P

    Journal: Nature

    Article Title: Pharmacological activation of REV-ERBs is lethal in cancer and oncogene induced senescence

    doi: 10.1038/nature25170

    Figure Lengend Snippet: REV-ERBs agonist SR9009 inhibits autophagy a–b , SR9009 treatment reduces the number of autophagosomes, as shown by immunofluorescence of LC3B, (n=biological independent samples MCF7 (n=6 mock), (n=5 SR9009) and T47D (n=5 mock) (n=4 SR9009) Mann–Whitney test one-tailed MCF7 20µM 24h * P =0.0152, T47D 20µM 48h ** P =0.0079; c–d SR9009 induces accumulation of p62 as shown by immunofluorescence; n= biological independent samples MCF7 (n=3 mock), (n=8 SR9009) and T47D (n=5 mock), (n=4 SR9009) Mann–Whitney test one-tailed 48h MCF7 p62 ** P =0.0061; 48h T47D ** P =0.0079; e, Inhibition of autophagy is confirmed by the immunoblot for p62 (20µM 48h, A375); f–g, Inhibition of autophagy precedes apoptosis induction as shown by immunofluorescence of p62, cleaved Caspase 3 and TUNEL assay; n=biological independent samples, Mann–Whitney test one-tailed, A375 20µM Cl. Casp. 3 48h (n=3) * P =0.0179; Cl. Casp. 3 72h (n=7) **** P

    Article Snippet: Apoptosis was evaluated by immunostaining of cleaved caspase 3 (Cell signaling #9664 1:200) and by TUNEL assay using In Situ Cell Death Detection Kit, Fluoresceine or TMR red (Roche).

    Techniques: Immunofluorescence, MANN-WHITNEY, One-tailed Test, Inhibition, TUNEL Assay

    Knockout of claudin-1 inhibits proliferation of cervical adenocarcinoma cells. (A-D) CLDN1 KO significantly inhibited proliferation of OMC4 cells in multiple assays, including cell count (A), colony formation assay (B), and immunostaining with anti–Ki-67 (proliferation marker) antibody (C). (D) Immunostaining with anti–cleaved caspase 3 (apoptosis marker) antibody. * P

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

    Article Title: Estrogen/GPR30 Signaling Contributes to the Malignant Potentials of ER-Negative Cervical Adenocarcinoma via Regulation of Claudin-1 Expression

    doi: 10.1016/j.neo.2018.08.010

    Figure Lengend Snippet: Knockout of claudin-1 inhibits proliferation of cervical adenocarcinoma cells. (A-D) CLDN1 KO significantly inhibited proliferation of OMC4 cells in multiple assays, including cell count (A), colony formation assay (B), and immunostaining with anti–Ki-67 (proliferation marker) antibody (C). (D) Immunostaining with anti–cleaved caspase 3 (apoptosis marker) antibody. * P

    Article Snippet: After incubation with a blocking buffer [0.01 mol/l PBS containing 5% bovine serum albumin (Sigma)], the sections were incubated with a rabbit polyclonal anti-Ki-67 (1:100, BioGenex) or anti-cleaved caspase-3 antibody (1:100, Cell Signaling Technology) at 4°C.

    Techniques: Knock-Out, Cell Counting, Colony Assay, Immunostaining, Marker

    Knockout of claudin-1 by the CRISPR-Cas9 system inhibits proliferation of cervical adenocarcinoma cells. (A) CLDN1 KO TMCC1 and OMC4 cells were established using the CRISPR-cas9 system. (B-E) CLDN1 KO significantly inhibited proliferation of TMCC1 cells in multiple assays including cell count (B), WST-8 assay (C), colony formation assay (D), and immunostaining with anti–Ki-67 (proliferation marker) antibody (E). (F) Immunostaining with anti–cleaved caspase 3 (apoptosis marker) antibody. (A) Western blotting. The relative intensity of each band is shown over each immunoblot after normalization for the levels of actin. * P

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

    Article Title: Estrogen/GPR30 Signaling Contributes to the Malignant Potentials of ER-Negative Cervical Adenocarcinoma via Regulation of Claudin-1 Expression

    doi: 10.1016/j.neo.2018.08.010

    Figure Lengend Snippet: Knockout of claudin-1 by the CRISPR-Cas9 system inhibits proliferation of cervical adenocarcinoma cells. (A) CLDN1 KO TMCC1 and OMC4 cells were established using the CRISPR-cas9 system. (B-E) CLDN1 KO significantly inhibited proliferation of TMCC1 cells in multiple assays including cell count (B), WST-8 assay (C), colony formation assay (D), and immunostaining with anti–Ki-67 (proliferation marker) antibody (E). (F) Immunostaining with anti–cleaved caspase 3 (apoptosis marker) antibody. (A) Western blotting. The relative intensity of each band is shown over each immunoblot after normalization for the levels of actin. * P

    Article Snippet: After incubation with a blocking buffer [0.01 mol/l PBS containing 5% bovine serum albumin (Sigma)], the sections were incubated with a rabbit polyclonal anti-Ki-67 (1:100, BioGenex) or anti-cleaved caspase-3 antibody (1:100, Cell Signaling Technology) at 4°C.

    Techniques: Knock-Out, CRISPR, Cell Counting, Colony Assay, Immunostaining, Marker, Western Blot

    Localization of ERS and UPR markers in the pregnant mouse uterus. Immunoflouresence studies were used to determine the localization of A) active CASP3 (Red) B) DDIT3 (Red) C) and HSPA5 (Green) at E 6, 8, 11, 13, 15, 17 and 19.

    Journal: PLoS ONE

    Article Title: Uterine Endoplasmic Reticulum Stress and Its Unfolded Protein Response May Regulate Caspase 3 Activation in the Pregnant Mouse Uterus

    doi: 10.1371/journal.pone.0075152

    Figure Lengend Snippet: Localization of ERS and UPR markers in the pregnant mouse uterus. Immunoflouresence studies were used to determine the localization of A) active CASP3 (Red) B) DDIT3 (Red) C) and HSPA5 (Green) at E 6, 8, 11, 13, 15, 17 and 19.

    Article Snippet: Tissue sections were then blocked in 1 X PBS containing 5% normal donkey serum and incubated in the primary antibody overnight at 4°C at the following concentrations DDIT3 1:100, cleaved CASP3 (Cell Signaling #9664 1:100), HSPA5 1:100.

    Techniques:

    Gestational profile of uterine CASP3 activation in the pregnant mouse uterus from E6–E19. A) Representative western blot of a gestational series of mouse uteri immunoblotted for cleaved (CL) and full length (FL) CASP3 protein (n = 3 for each time point). Relative optical density (ROD) of B) cleaved and C) full length CASP3 normalized to the loading control, PDI. Data are represented as Mean ROD ± SE. N = 3 for each time point. Data labeled with different letters are significantly different from each other (P

    Journal: PLoS ONE

    Article Title: Uterine Endoplasmic Reticulum Stress and Its Unfolded Protein Response May Regulate Caspase 3 Activation in the Pregnant Mouse Uterus

    doi: 10.1371/journal.pone.0075152

    Figure Lengend Snippet: Gestational profile of uterine CASP3 activation in the pregnant mouse uterus from E6–E19. A) Representative western blot of a gestational series of mouse uteri immunoblotted for cleaved (CL) and full length (FL) CASP3 protein (n = 3 for each time point). Relative optical density (ROD) of B) cleaved and C) full length CASP3 normalized to the loading control, PDI. Data are represented as Mean ROD ± SE. N = 3 for each time point. Data labeled with different letters are significantly different from each other (P

    Article Snippet: Tissue sections were then blocked in 1 X PBS containing 5% normal donkey serum and incubated in the primary antibody overnight at 4°C at the following concentrations DDIT3 1:100, cleaved CASP3 (Cell Signaling #9664 1:100), HSPA5 1:100.

    Techniques: Activation Assay, Western Blot, Labeling

    A model representing the potential role of ERSR and the UPR in the gestational regulation of uterine CASP3 activation and inhibition.

    Journal: PLoS ONE

    Article Title: Uterine Endoplasmic Reticulum Stress and Its Unfolded Protein Response May Regulate Caspase 3 Activation in the Pregnant Mouse Uterus

    doi: 10.1371/journal.pone.0075152

    Figure Lengend Snippet: A model representing the potential role of ERSR and the UPR in the gestational regulation of uterine CASP3 activation and inhibition.

    Article Snippet: Tissue sections were then blocked in 1 X PBS containing 5% normal donkey serum and incubated in the primary antibody overnight at 4°C at the following concentrations DDIT3 1:100, cleaved CASP3 (Cell Signaling #9664 1:100), HSPA5 1:100.

    Techniques: Activation Assay, Inhibition

    Mangafodipir rescues follicles from anticancer drug-induced apoptosis. a Representative images of immunohistochemistry for cleaved caspase-3 in ovaries treated with 7.5 (CIS7.5) or 20 (CIS20) mg/kg cisplatin, or 7.5 (PTX7.5) or 20 (PTX20) mg/kg paclitaxel, with or without mangafodipir (+/− Mangafodipir, respectively). Bar, 100 μm. b Cleaved caspase-3 (CC3) immunoreactive intensity scores. Mean ± SD, *, p

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: Protective effects of mangafodipir against chemotherapy-induced ovarian damage in mice

    doi: 10.1186/s12958-018-0426-y

    Figure Lengend Snippet: Mangafodipir rescues follicles from anticancer drug-induced apoptosis. a Representative images of immunohistochemistry for cleaved caspase-3 in ovaries treated with 7.5 (CIS7.5) or 20 (CIS20) mg/kg cisplatin, or 7.5 (PTX7.5) or 20 (PTX20) mg/kg paclitaxel, with or without mangafodipir (+/− Mangafodipir, respectively). Bar, 100 μm. b Cleaved caspase-3 (CC3) immunoreactive intensity scores. Mean ± SD, *, p

    Article Snippet: Next, sections were reacted with rabbit anti-cleaved caspase-3 antibody (1:100; Cell Signaling Technology Japan, K.K., Tokyo, Japan), rabbit anti-4-HNE antibody (1:1500; ab46545; Abcam, Tokyo, Japan) and rabbit anti-Ki67 antibody (1:500; ab92742; Abcam, Cambridge, UK) overnight at 4 °C.

    Techniques: Immunohistochemistry

    Evaluation of V-9302  in vivo  in HCC1806 cell line xenograft-bearing mice ( A ) Tumor volumetric analysis of mice treated with vehicle or V-9302 (75 mg/kg per day) for 10 days.  n  = 5 mice per group. Treatment started 9 days post injection. P value at day 10 determined by Student’s  t  test. ( B ) Immunofluorescence analysis of tumor tissues harvested from vehicle- or V-9302-treated mice; LC3B and pAKT (Ser473) shown (pink). CD-31 positive vessels shown in green and nuclei in blue (DAPI). 20× magnification. P values determined by Student’s  t  test. ( C ) Effects of V-9302 treatment on pS6-positive cells and caspase 3-positive cells by immunohistochemistry. Quantitative analysis consisted of the mean counts of at least three representative fields from three vehicle and three V-9302-treated mice. For box plots, center line is plotted at the median; the box spans from the first quartile to the third quartile; whiskers represent min to max. Error bars represent ± std. dev.

    Journal: Nature medicine

    Article Title: Pharmacological Blockade of ASCT2-dependent Glutamine Transport Leads To Anti-tumor Efficacy in Preclinical Models

    doi: 10.1038/nm.4464

    Figure Lengend Snippet: Evaluation of V-9302 in vivo in HCC1806 cell line xenograft-bearing mice ( A ) Tumor volumetric analysis of mice treated with vehicle or V-9302 (75 mg/kg per day) for 10 days. n = 5 mice per group. Treatment started 9 days post injection. P value at day 10 determined by Student’s t test. ( B ) Immunofluorescence analysis of tumor tissues harvested from vehicle- or V-9302-treated mice; LC3B and pAKT (Ser473) shown (pink). CD-31 positive vessels shown in green and nuclei in blue (DAPI). 20× magnification. P values determined by Student’s t test. ( C ) Effects of V-9302 treatment on pS6-positive cells and caspase 3-positive cells by immunohistochemistry. Quantitative analysis consisted of the mean counts of at least three representative fields from three vehicle and three V-9302-treated mice. For box plots, center line is plotted at the median; the box spans from the first quartile to the third quartile; whiskers represent min to max. Error bars represent ± std. dev.

    Article Snippet: Tissues were sectioned (5 µm thickness) and stained for pS6 (Cell Signaling, #4858) and caspase 3 (Cell Signaling #9661).

    Techniques: In Vivo, Mouse Assay, Injection, Immunofluorescence, Immunohistochemistry

    Evaluation of V-9302 in vivo ( A ) Pharmacodynamic [ 18 F]-4F-Gln PET imaging prior to and 4 h following a single administration of V-9302 (75 mg/kg) in HCC1806 cell line xenograft-bearing mice (arrows indicate xenograft tumor on right flank*). ( B ) Mean time activity curves (TACs) from tumor regions of interest ( n = 4 measurements per condition); data prior to and 4 hrs following V-9302 administration. ( C ) P values determined by Student’s t test. Quantified tracer accumulation in xenograft tumors, muscle, and liver ( n = 4 measurements per condition). Volumetric analysis over 21 day treatment regimen (Vehicle or V-9302; 75 mg/kg, daily) of HCT-116 ( D ) and HT29 ( F ) cell line xenografts propagated in athymic nude mice ( n = 10 mice per group). Treatment started 12 days post tumor injection for HCT-116 and 4 days post injection for HT29. P values on day 21 determined by Student’s t test. Immunohistochemistry for pS6 and caspase 3 in vehicle-treated or V-9302-treated HCT-116 ( E ) and HT29 ( G ) xenografts. Representative photomicrographs and quantitation shown; magnification 20×. P values determined by Student’s t test. ( H ) Volumetric analysis over 31 day treatment regimen (Vehicle or V-9302; 75 mg/kg, daily) on athymic nude mice bearing patient-derived xenograft tumors (PDX A 008, F3 generation, treatment started 28 days post implantation, KRAS G12V ;p53 R248Q ;PTEN L140Y ; n = 10 mice per group) P value on day 31 determined by Student’s t test. ( I ) Photographs of A 008 PDX-bearing mice treated with V-9302 or vehicle; day 16 of 31. (Error bars represent ± std. dev. *Central photopenia observed.

    Journal: Nature medicine

    Article Title: Pharmacological Blockade of ASCT2-dependent Glutamine Transport Leads To Anti-tumor Efficacy in Preclinical Models

    doi: 10.1038/nm.4464

    Figure Lengend Snippet: Evaluation of V-9302 in vivo ( A ) Pharmacodynamic [ 18 F]-4F-Gln PET imaging prior to and 4 h following a single administration of V-9302 (75 mg/kg) in HCC1806 cell line xenograft-bearing mice (arrows indicate xenograft tumor on right flank*). ( B ) Mean time activity curves (TACs) from tumor regions of interest ( n = 4 measurements per condition); data prior to and 4 hrs following V-9302 administration. ( C ) P values determined by Student’s t test. Quantified tracer accumulation in xenograft tumors, muscle, and liver ( n = 4 measurements per condition). Volumetric analysis over 21 day treatment regimen (Vehicle or V-9302; 75 mg/kg, daily) of HCT-116 ( D ) and HT29 ( F ) cell line xenografts propagated in athymic nude mice ( n = 10 mice per group). Treatment started 12 days post tumor injection for HCT-116 and 4 days post injection for HT29. P values on day 21 determined by Student’s t test. Immunohistochemistry for pS6 and caspase 3 in vehicle-treated or V-9302-treated HCT-116 ( E ) and HT29 ( G ) xenografts. Representative photomicrographs and quantitation shown; magnification 20×. P values determined by Student’s t test. ( H ) Volumetric analysis over 31 day treatment regimen (Vehicle or V-9302; 75 mg/kg, daily) on athymic nude mice bearing patient-derived xenograft tumors (PDX A 008, F3 generation, treatment started 28 days post implantation, KRAS G12V ;p53 R248Q ;PTEN L140Y ; n = 10 mice per group) P value on day 31 determined by Student’s t test. ( I ) Photographs of A 008 PDX-bearing mice treated with V-9302 or vehicle; day 16 of 31. (Error bars represent ± std. dev. *Central photopenia observed.

    Article Snippet: Tissues were sectioned (5 µm thickness) and stained for pS6 (Cell Signaling, #4858) and caspase 3 (Cell Signaling #9661).

    Techniques: In Vivo, Positron Emission Tomography, Imaging, Mouse Assay, Activity Assay, Injection, Immunohistochemistry, Quantitation Assay, Derivative Assay

    Haematopoietic and gastrointestinal failure in AIMP3 mKO mouse. ( A ) Hypotrophy of haematopoietic and intestinal organs in AIMP3 mKO. At day 30, the thymus, spleen, bone marrow and intestine from AIMP3 CON (left column) and AIMP3 mKO (right column) were isolated, and tissue sections were stained with haematoxylin-eosin (HE). The inset shows a low-resolution image of the HE-stained tissue. Dotted lines mark the boundary between the cortex (C) and medulla (M) in the thymic lobe and between the white pulp (WP) and red pulp (RP) in the spleen, respectively. Arrows and arrowheads in the intestine indicate elongated crypts and blunted villi, respectively, in AIMP3 mKO mice. ( B ) Bone marrow isolated from AIMP3 CON and AIMP3 mKO mice were immunostained with antibodies against cleaved caspase-3. ( C ) Splenocytes from AIMP3 CON (left) and AIMP3 mKO (right) mice were analysed by FACS cytometry using the markers CD3ε (pan-T cells) and CD45R (pan-B cells). The number indicates the percentage of the cells in the quadrant. Representative results from three animals are shown. ( D ) CD3ε–(left), CD45R- (middle) and CD11b- (right) positive cells were analysed by FACS in spleen and bone marrow from AIMP3 CON and AIMP3 mKO mice. Three to five animals per group were used. Error bars indicate standard error. Asterisks denote significant difference between groups by Student’s t-test (*p

    Journal: Scientific Reports

    Article Title: AIMP3 Deletion Induces Acute Radiation Syndrome-like Phenotype in Mice

    doi: 10.1038/s41598-018-33303-3

    Figure Lengend Snippet: Haematopoietic and gastrointestinal failure in AIMP3 mKO mouse. ( A ) Hypotrophy of haematopoietic and intestinal organs in AIMP3 mKO. At day 30, the thymus, spleen, bone marrow and intestine from AIMP3 CON (left column) and AIMP3 mKO (right column) were isolated, and tissue sections were stained with haematoxylin-eosin (HE). The inset shows a low-resolution image of the HE-stained tissue. Dotted lines mark the boundary between the cortex (C) and medulla (M) in the thymic lobe and between the white pulp (WP) and red pulp (RP) in the spleen, respectively. Arrows and arrowheads in the intestine indicate elongated crypts and blunted villi, respectively, in AIMP3 mKO mice. ( B ) Bone marrow isolated from AIMP3 CON and AIMP3 mKO mice were immunostained with antibodies against cleaved caspase-3. ( C ) Splenocytes from AIMP3 CON (left) and AIMP3 mKO (right) mice were analysed by FACS cytometry using the markers CD3ε (pan-T cells) and CD45R (pan-B cells). The number indicates the percentage of the cells in the quadrant. Representative results from three animals are shown. ( D ) CD3ε–(left), CD45R- (middle) and CD11b- (right) positive cells were analysed by FACS in spleen and bone marrow from AIMP3 CON and AIMP3 mKO mice. Three to five animals per group were used. Error bars indicate standard error. Asterisks denote significant difference between groups by Student’s t-test (*p

    Article Snippet: Cleaved caspase-3 antibody (Cell signalling #9661, 1:200) and Dako system for rabbit was used for colorimetric development.

    Techniques: Isolation, Staining, Mouse Assay, FACS, Cytometry

    AgNP-induced occurrence of systemic apoptosis. Immunofluorescent signals of the apoptotic biomarker “active caspase 3” in multiple tissues (brain, salivary gland, wing disc and gut) dissected from AgNP-exposed 3 rd -instar wandering larvae.

    Journal: Scientific Reports

    Article Title: Silver nanoparticles have lethal and sublethal adverse effects on development and longevity by inducing ROS-mediated stress responses

    doi: 10.1038/s41598-018-20728-z

    Figure Lengend Snippet: AgNP-induced occurrence of systemic apoptosis. Immunofluorescent signals of the apoptotic biomarker “active caspase 3” in multiple tissues (brain, salivary gland, wing disc and gut) dissected from AgNP-exposed 3 rd -instar wandering larvae.

    Article Snippet: The rabbit anti-cleaved caspase 3 (1:250, Cell Signaling Technology, MA, USA) and rabbit anti-phosphorylated H2AX (1:500, Rockland, PA, USA) primary antibodies were used to quantitate the levels of AgNP-induced apoptosis and DNA damage (double-stranded breaks), respectively.

    Techniques: Biomarker Assay

    APE1/Ref-1 redox inhibition induces G1 cell arrest ( A ) PC-3 and C4-2 cell lines were treated with DMSO or APX2009 (9 and 14 µM, respectively) for 48 hours. Representative images were taken at 20× Magnification. Scale bar = 50 µm. ( B ) Immunoblotting was performed and membranes were probed with antibodies for Cleaved Caspase 3, Total Caspase, Cyclin B1, Cdc2, survivin and Actin as labeled. ( C ) PC-3 and C4-2 cells were treated with DMSO or APX2009 (9 and 14 µM, respectively) for 48 hrs and then collected and stained with RNAse/PI wash. Flow Cytometry was then performed.  n  =3,  * -denoting  p

    Journal: Oncotarget

    Article Title: APE1/Ref-1 redox-specific inhibition decreases survivin protein levels and induces cell cycle arrest in prostate cancer cells

    doi: 10.18632/oncotarget.23493

    Figure Lengend Snippet: APE1/Ref-1 redox inhibition induces G1 cell arrest ( A ) PC-3 and C4-2 cell lines were treated with DMSO or APX2009 (9 and 14 µM, respectively) for 48 hours. Representative images were taken at 20× Magnification. Scale bar = 50 µm. ( B ) Immunoblotting was performed and membranes were probed with antibodies for Cleaved Caspase 3, Total Caspase, Cyclin B1, Cdc2, survivin and Actin as labeled. ( C ) PC-3 and C4-2 cells were treated with DMSO or APX2009 (9 and 14 µM, respectively) for 48 hrs and then collected and stained with RNAse/PI wash. Flow Cytometry was then performed. n =3, * -denoting p

    Article Snippet: Proteins were transferred to polyvinylidene difluoride (PVDF) membranes, blocked for 24 hours [(10% Dry milk, 5% BSA, .05% NaN3 ) in 1xPBS( 2.7 mM KCl, 1.5 mM KH2 PO4 , 136 mM NaCl, 8 mM Na2 HPO4 )-Tween 20] and incubated overnight with one of the following primary antibodies: mouse β-actin (1:2500, ThermoFisher Scientific), mouse APE1/Ref-1 (1:1000, Novus Biologicals), rabbit survivin (1:500, Cell Signaling Technologies), rabbit Bcl-2 (1:500, Cell Signaling Technologies), rabbit Mcl-1 (1:500, Cell Signaling Technologies), rabbit Cleaved Caspase 3 (1:250, Cell Signaling Technologies), rabbit Total Caspase 3 (1:1000, Cell Signaling Technologies), rabbit Cyclin B1 (1:500, Cell Signaling Technologies), Cdc2 (1:1000, Cell Signaling Technologies) and rabbit GAPDH (1:1000, Cell Signaling Technologies).

    Techniques: Inhibition, Labeling, Staining, Flow Cytometry, Cytometry

    p53 25,26,53,54/+  Embryos Exhibit Additional Features of CHARGE Syndrome and p53-Dependent Cellular Responses (a)  Double outlet right ventricle (DORV) in E13.5 p53 25,26,53,54/+  heart (50%, n=6). Top: Main pulmonary artery (MPA) connects via pulmonary valve (PV) to right ventricle (RV) in both control and p53 25,26,53,54/+  embryo. Bottom: Aorta (Ao) in control embryo connects to left ventricle (LV) via aortic valve (AV) Φ . Aorta in p53 25,26,53,54/+  embryo connects to RV via AV*. (b)  Abnormal atrioventricular cushions in E13.5 p53 25,26,53,54/+  heart (75%, n=4) fail to elongateinto mature mitral (mv, arrowhead) and tricuspid (tv, arrow) valves. RA: right atrium; LA: left atrium.  (c)  E13.5  p53 25,26,53,54/+  kidneys are smaller (79%), with fewer average glomeruli (13 vs. 3; n=5; arrows), than controls. (d) p53 25,26,53,54/+  embryonic phenotypes observed in CHARGE (+present, −absent).  (e)  Left: Cleaved-caspase 3 (CC3; Top) and p53 (Bottom) immunohistochemistry in E15.5 retinas. Arrows: CC3-positive cells. Right: CC3-positive cells per retinal area. ***p-value=0.007; one-tailed Welsh’s t-test (n=5).  (f)  BrdU immunofluorescence in E9.5 Pax3 +  NCCs (delineated by green-dotted line;  Extended-Data Fig. 6c ). Right: Percentage BrdU-positive cells per total Pax3 +  NCCs ***p-value=0.004 one-tailed Student’s t-test (n=4).

    Journal: Nature

    Article Title: Inappropriate p53 Activation During Development Induces Features of CHARGE Syndrome

    doi: 10.1038/nature13585

    Figure Lengend Snippet: p53 25,26,53,54/+ Embryos Exhibit Additional Features of CHARGE Syndrome and p53-Dependent Cellular Responses (a) Double outlet right ventricle (DORV) in E13.5 p53 25,26,53,54/+ heart (50%, n=6). Top: Main pulmonary artery (MPA) connects via pulmonary valve (PV) to right ventricle (RV) in both control and p53 25,26,53,54/+ embryo. Bottom: Aorta (Ao) in control embryo connects to left ventricle (LV) via aortic valve (AV) Φ . Aorta in p53 25,26,53,54/+ embryo connects to RV via AV*. (b) Abnormal atrioventricular cushions in E13.5 p53 25,26,53,54/+ heart (75%, n=4) fail to elongateinto mature mitral (mv, arrowhead) and tricuspid (tv, arrow) valves. RA: right atrium; LA: left atrium. (c) E13.5 p53 25,26,53,54/+ kidneys are smaller (79%), with fewer average glomeruli (13 vs. 3; n=5; arrows), than controls. (d) p53 25,26,53,54/+ embryonic phenotypes observed in CHARGE (+present, −absent). (e) Left: Cleaved-caspase 3 (CC3; Top) and p53 (Bottom) immunohistochemistry in E15.5 retinas. Arrows: CC3-positive cells. Right: CC3-positive cells per retinal area. ***p-value=0.007; one-tailed Welsh’s t-test (n=5). (f) BrdU immunofluorescence in E9.5 Pax3 + NCCs (delineated by green-dotted line; Extended-Data Fig. 6c ). Right: Percentage BrdU-positive cells per total Pax3 + NCCs ***p-value=0.004 one-tailed Student’s t-test (n=4).

    Article Snippet: Whole-mount cleaved-caspase 3 staining was performed as described with anti-cleaved-caspase 3 antibody (Cell Signaling #9664) and developed with DAB (Vector Labs).

    Techniques: Immunohistochemistry, One-tailed Test, Immunofluorescence

    p53 25,26,53,54/+  Embryo Tissues Display Increased Apoptosis and Decreased Proliferation (a)  Left: Immunofluorescence for Phospho-Histone H3 (red) in the retina of E13.5 control and  p53 25,26,53,54/+ embryos. Right: Quantification of Phospho-Histone H3 positive cells per retina area relative to littermate controls. **p-value=0.006 by one-tailed Welsh’s t-test (n=4). ( b ) Left: Immunohistochemistry for cleaved-caspase 3 (CC3) in thymi of control (left) and  p53 25,26,53,54/+  (right) embryos. Inset: close-up image of cleaved-caspase 3 positive region. Right: Quantification of CC3-positive cells per thymic area. *p-value=0.02 by one-tailed Student’s t-test (n=4).  (c)  Immunofluorescence for Pax3 (green) in neural crest cells of E9.5 control and p53 25,26,53,54/+  embryos was used to identify neural crest cells in   Figure 2f .  (d) Left: Immunofluorescence for cleaved-caspase 3 (CC3, red) and Pax3 (green) in neural crest cells of E9.5 control and  p53 25,26,53,54/+ embryos.  p53 25,26,53,54/+  embryos have more apoptotic (red) neural crest cells, as determined by Pax3-positive staining (green), compared to control littermates. Right: Quantification of CC3 positive cells per total neural crest cell number. p-value=0.14 by one-tailed Student’s t-test (n=4). (e)  Left: Immunofluorescence for cleaved-caspase 3 (CC3, red) in otic vesicle of E9.5 control and  p53 25,26,53,54/+  embryos. Right: Quantification of CC3 positive cells per total cell number. *p-value=0.03 by one-tailed Student’s t-test (n=3). ( f ) Whole-mount cleaved-caspase 3 staining in E8.5 control and p53 25,26,53,54/+  embryos reveals enhanced apoptosis in the neuroepithelium of  p53 25,26,53,54/+  embryos (right) but not in controls (left). Close-up shows magnification of the caudal neuroepithelium (bottom). Arrows indicate cleaved-caspase 3 positive regions.

    Journal: Nature

    Article Title: Inappropriate p53 Activation During Development Induces Features of CHARGE Syndrome

    doi: 10.1038/nature13585

    Figure Lengend Snippet: p53 25,26,53,54/+ Embryo Tissues Display Increased Apoptosis and Decreased Proliferation (a) Left: Immunofluorescence for Phospho-Histone H3 (red) in the retina of E13.5 control and p53 25,26,53,54/+ embryos. Right: Quantification of Phospho-Histone H3 positive cells per retina area relative to littermate controls. **p-value=0.006 by one-tailed Welsh’s t-test (n=4). ( b ) Left: Immunohistochemistry for cleaved-caspase 3 (CC3) in thymi of control (left) and p53 25,26,53,54/+ (right) embryos. Inset: close-up image of cleaved-caspase 3 positive region. Right: Quantification of CC3-positive cells per thymic area. *p-value=0.02 by one-tailed Student’s t-test (n=4). (c) Immunofluorescence for Pax3 (green) in neural crest cells of E9.5 control and p53 25,26,53,54/+ embryos was used to identify neural crest cells in Figure 2f . (d) Left: Immunofluorescence for cleaved-caspase 3 (CC3, red) and Pax3 (green) in neural crest cells of E9.5 control and p53 25,26,53,54/+ embryos. p53 25,26,53,54/+ embryos have more apoptotic (red) neural crest cells, as determined by Pax3-positive staining (green), compared to control littermates. Right: Quantification of CC3 positive cells per total neural crest cell number. p-value=0.14 by one-tailed Student’s t-test (n=4). (e) Left: Immunofluorescence for cleaved-caspase 3 (CC3, red) in otic vesicle of E9.5 control and p53 25,26,53,54/+ embryos. Right: Quantification of CC3 positive cells per total cell number. *p-value=0.03 by one-tailed Student’s t-test (n=3). ( f ) Whole-mount cleaved-caspase 3 staining in E8.5 control and p53 25,26,53,54/+ embryos reveals enhanced apoptosis in the neuroepithelium of p53 25,26,53,54/+ embryos (right) but not in controls (left). Close-up shows magnification of the caudal neuroepithelium (bottom). Arrows indicate cleaved-caspase 3 positive regions.

    Article Snippet: Whole-mount cleaved-caspase 3 staining was performed as described with anti-cleaved-caspase 3 antibody (Cell Signaling #9664) and developed with DAB (Vector Labs).

    Techniques: Immunofluorescence, One-tailed Test, Immunohistochemistry, Staining