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Knockdown of LEF1 induces apoptosis in PAAD cells. A–C, Flow cytometry detection of apoptosis and apoptosis rate statistics: apoptosis rate was higher in AsPC-1/shLEF1 and BxPC-3/shLEF1. D–F, Calcein AM/PI staining assay results and statistics: the live cells decreased while the dead cells increased in AsPC-1/shLEF1 and BxPC-3/shLEF1. G, ATP production assay: the ATP production decreased in AsPC-1/shLEF1 and BxPC-3/shLEF1. H, LDH release assay showed an increase in LDH release in AsPC-1/shLEF1 and BxPC-3/shLEF1. I–M, WB results and quantitative analysis: the level of Bcl-2 protein decreased in AsPC-1/shLEF1 and BxPC-3/shLEF1, while BAX, Cleaved caspase-3 and <t>Cleaved</t> <t>caspase-9</t> increased (n = 3, * P <0.05, ** P <0.01).
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Knockdown of LEF1 induces apoptosis in PAAD cells. A–C, Flow cytometry detection of apoptosis and apoptosis rate statistics: apoptosis rate was higher in AsPC-1/shLEF1 and BxPC-3/shLEF1. D–F, Calcein AM/PI staining assay results and statistics: the live cells decreased while the dead cells increased in AsPC-1/shLEF1 and BxPC-3/shLEF1. G, ATP production assay: the ATP production decreased in AsPC-1/shLEF1 and BxPC-3/shLEF1. H, LDH release assay showed an increase in LDH release in AsPC-1/shLEF1 and BxPC-3/shLEF1. I–M, WB results and quantitative analysis: the level of Bcl-2 protein decreased in AsPC-1/shLEF1 and BxPC-3/shLEF1, while BAX, Cleaved caspase-3 and <t>Cleaved</t> <t>caspase-9</t> increased (n = 3, * P <0.05, ** P <0.01).
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Knockdown of LEF1 induces apoptosis in PAAD cells. A–C, Flow cytometry detection of apoptosis and apoptosis rate statistics: apoptosis rate was higher in AsPC-1/shLEF1 and BxPC-3/shLEF1. D–F, Calcein AM/PI staining assay results and statistics: the live cells decreased while the dead cells increased in AsPC-1/shLEF1 and BxPC-3/shLEF1. G, ATP production assay: the ATP production decreased in AsPC-1/shLEF1 and BxPC-3/shLEF1. H, LDH release assay showed an increase in LDH release in AsPC-1/shLEF1 and BxPC-3/shLEF1. I–M, WB results and quantitative analysis: the level of Bcl-2 protein decreased in AsPC-1/shLEF1 and BxPC-3/shLEF1, while BAX, Cleaved caspase-3 and <t>Cleaved</t> <t>caspase-9</t> increased (n = 3, * P <0.05, ** P <0.01).
Caspase 9, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Knockdown of LEF1 induces apoptosis in PAAD cells. A–C, Flow cytometry detection of apoptosis and apoptosis rate statistics: apoptosis rate was higher in AsPC-1/shLEF1 and BxPC-3/shLEF1. D–F, Calcein AM/PI staining assay results and statistics: the live cells decreased while the dead cells increased in AsPC-1/shLEF1 and BxPC-3/shLEF1. G, ATP production assay: the ATP production decreased in AsPC-1/shLEF1 and BxPC-3/shLEF1. H, LDH release assay showed an increase in LDH release in AsPC-1/shLEF1 and BxPC-3/shLEF1. I–M, WB results and quantitative analysis: the level of Bcl-2 protein decreased in AsPC-1/shLEF1 and BxPC-3/shLEF1, while BAX, Cleaved caspase-3 and <t>Cleaved</t> <t>caspase-9</t> increased (n = 3, * P <0.05, ** P <0.01).
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Knockdown of LEF1 induces apoptosis in PAAD cells. A–C, Flow cytometry detection of apoptosis and apoptosis rate statistics: apoptosis rate was higher in AsPC-1/shLEF1 and BxPC-3/shLEF1. D–F, Calcein AM/PI staining assay results and statistics: the live cells decreased while the dead cells increased in AsPC-1/shLEF1 and BxPC-3/shLEF1. G, ATP production assay: the ATP production decreased in AsPC-1/shLEF1 and BxPC-3/shLEF1. H, LDH release assay showed an increase in LDH release in AsPC-1/shLEF1 and BxPC-3/shLEF1. I–M, WB results and quantitative analysis: the level of Bcl-2 protein decreased in AsPC-1/shLEF1 and BxPC-3/shLEF1, while BAX, Cleaved caspase-3 and Cleaved caspase-9 increased (n = 3, * P <0.05, ** P <0.01).

Journal: Pancreas

Article Title: Knockdown of Lymphoid Enhancer-binding Factor 1 Inhibits Pancreatic Adenocarcinoma Growth and Neoangiogenesis by Curbing Notch1 and Nuclear Factor Kappa B Signaling Pathways

doi: 10.1097/MPA.0000000000002562

Figure Lengend Snippet: Knockdown of LEF1 induces apoptosis in PAAD cells. A–C, Flow cytometry detection of apoptosis and apoptosis rate statistics: apoptosis rate was higher in AsPC-1/shLEF1 and BxPC-3/shLEF1. D–F, Calcein AM/PI staining assay results and statistics: the live cells decreased while the dead cells increased in AsPC-1/shLEF1 and BxPC-3/shLEF1. G, ATP production assay: the ATP production decreased in AsPC-1/shLEF1 and BxPC-3/shLEF1. H, LDH release assay showed an increase in LDH release in AsPC-1/shLEF1 and BxPC-3/shLEF1. I–M, WB results and quantitative analysis: the level of Bcl-2 protein decreased in AsPC-1/shLEF1 and BxPC-3/shLEF1, while BAX, Cleaved caspase-3 and Cleaved caspase-9 increased (n = 3, * P <0.05, ** P <0.01).

Article Snippet: The first antibodies were as follows: LEF1 (ab137872, 1:1000, Abcam), proliferating cell nuclear antigen (PCNA, ab29, 1:1000, Abcam), MMP-2 (4022S, 1:1000, CST), MMP-9 (3852S, 1:1000, CST), BAX (2772S, 1:1000, CST), Bcl-2 (15071S, 1:1000, CST), Cleaved caspase-3 (9661S, 1:1000, CST), Cleaved caspase-9 (9509S, 1:1000, CST), vascular endothelial growth factor-A (VEGFA, ab214424, 1:1000, Abcam), total Notch1 (ab52627, 1:1000, Abcam), NICD (4147, 1:1000, CST), Hes1 (ab71559, 1:1000, Abcam), Hey1 (ab235173, 1:1000, Abcam), Jagged1 (ab300561, 1:1000, Abcam), P65 (ab32536, 1:1000, Abcam), p-P65 (3033S, 1:1000, CST), GAPDH (5174S, 1:5000, CST), HRP-anti-Rabbit (A0362, 1:1000, Beyotime), HRP-anti-Mouse (A0350, 1:1000, Beyotime).

Techniques: Knockdown, Flow Cytometry, Staining, Lactate Dehydrogenase Assay