Journal: Cell Death and Differentiation

Article Title: Mcl-1 and Bcl-xL are essential for survival of the developing nervous system

doi: 10.1038/s41418-018-0225-1

Figure Lengend Snippet: Conditional deletion of both mcl-1 and bcl-x—a double conditional knockout (DKO) results in complete loss of the CNS by E14. a Photomicrographs of representative cresyl violet-stained sagittal sections of E14 CTL, MKO, BKO, and DKO embryos. Representative higher magnification photomicrographs of cresyl violet stained sections of (b) cortex and (c) spinal cords for each genotype. Scale bars = 2000 μm (a), 100 μm (b, c)

Article Snippet: Primary antibodies Mcl-1 (Rockland, 600-401-394, RRID:AB_2266446), Bcl-xL (Cell Signaling Technology, Cat# 2764, RRID:AB_2228008), and β-actin (Sigma-Aldrich, Cat# A5316, RRID:AB_476743) were applied to blots overnight, followed by the application of a horse-radish peroxidase-tagged secondary antibody (Bio-Rad, 1706515/1706516, Mississauga, Ontario, Canada).

Techniques: Knock-Out, Staining

Journal: Cell Death and Differentiation

Article Title: Mcl-1 and Bcl-xL are essential for survival of the developing nervous system

doi: 10.1038/s41418-018-0225-1

Figure Lengend Snippet: Gene dosage affects the onset of apoptosis in the spinal cord. a Photomicrograph of a cresyl violet stained sagittal section through an E11 mouse embryo. Black box indicates region of spinal cord examined. b Representative photomicrographs of sagittal spinal cord sections from E11 CTL, Mcl-1 HET:Bcl-x HET, MKO, MKO:Bcl-x HET, BKO, Mcl-1 HET:BKO, and DKO animals (n = a minimum of 3/genotype). Apoptotic cells are labeled with active caspase-3 (AC3) immunohistochemistry, while cresyl violet staining shows tissue histology. Larger boxed area in b indicates region of higher magnification photomicrographs for active caspase-3 stained sections, while smaller inset indicates region shown in cresyl violet photomicrographs. R rostral, C caudal, D dorsal, V ventral

Article Snippet: Primary antibodies Mcl-1 (Rockland, 600-401-394, RRID:AB_2266446), Bcl-xL (Cell Signaling Technology, Cat# 2764, RRID:AB_2228008), and β-actin (Sigma-Aldrich, Cat# A5316, RRID:AB_476743) were applied to blots overnight, followed by the application of a horse-radish peroxidase-tagged secondary antibody (Bio-Rad, 1706515/1706516, Mississauga, Ontario, Canada).

Techniques: Staining, Labeling, Immunohistochemistry

Journal: Cell Death and Differentiation

Article Title: Mcl-1 and Bcl-xL are essential for survival of the developing nervous system

doi: 10.1038/s41418-018-0225-1

Figure Lengend Snippet: An anti-apoptotic role for Bcl-xL in neural precursor cell populations. a Representative coronal spinal cord sections from E11 Mcl-1 HET:Bcl-x HET, MKO, MKO:Bcl-x HET, BKO, and Mcl-1 HET:BKO embryos immunostained with antibodies to AC3 and Ki67. Higher magnification of boxed areas in a showing double labeled cells. b Representative coronal spinal cord sections from E11 Mcl-1 HET:Bcl-x HET (TRH triple heterozygous), MKO, MKO:Bcl-x HET, BKO, and Mcl-1 HET:BKO embryos immunostained with antibodies to AC3 and βIII-tubulin (Tuj1). Higher magnification of boxed areas in b showing double-labeled cells. c Counts of apoptotic cells within the proliferating population of the spinal cord as shown in a. There was no significant differences in the number of apoptotic cells between the TRH, BKO, and BKO:Mcl-1 HET. In contrast, the MKO:Bcl-x HET had significantly more apoptotic cells than the MKO (p < 0.01) and the TRH, BKO, and BKO:Mcl-1 HET (p < 0.001) and the MKO had significantly more apoptotic cells than the BKO:Mcl-1 HET (p < 0.05) and the TRH, BKO, MKO:Bcl-x HET (p < 0.01). d Counts of apoptotic cells within the ventral lateral horns of the spinal cord as shown in b. Overall fewer apoptotic cells were observed in the ventral horns versus in the proliferating NPCs in the center of the spinal cord. A significant increase in the number of apoptotic cells was observed in the BKO:Mcl-1 HET versus the TRH (p < 0.05). Individual means ± SD are shown for each embryo. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: Primary antibodies Mcl-1 (Rockland, 600-401-394, RRID:AB_2266446), Bcl-xL (Cell Signaling Technology, Cat# 2764, RRID:AB_2228008), and β-actin (Sigma-Aldrich, Cat# A5316, RRID:AB_476743) were applied to blots overnight, followed by the application of a horse-radish peroxidase-tagged secondary antibody (Bio-Rad, 1706515/1706516, Mississauga, Ontario, Canada).

Techniques: Labeling

Journal: Cell Death and Differentiation

Article Title: Mcl-1 and Bcl-xL are essential for survival of the developing nervous system

doi: 10.1038/s41418-018-0225-1

Figure Lengend Snippet: Deletion of Bax rescues cell death in the MKO and BKO embryo. a Representative horizontal sections through the E11 lumbar spinal cord of Mcl-1 HET:Bax HET, MKO, and MKO:Bax Null embryos immunostained with antibodies to AC3 showed that deletion of Bax rescues the majority of apoptotic cells in the MKO spinal cord. b Higher magnification of the area for apoptotic cell counts (AC3) and total nuclei counts (Hoechst) in each of the genotypes. c Representative coronal sections through the spinal cord of E13 Bcl-x HET:Bax Het, BKO, and BKO:Bax Null embryos. d Higher magnification of the area for apoptotic cell counts (AC3) and total nuclei counts (Hoechst) in each of the genotypes

Article Snippet: Primary antibodies Mcl-1 (Rockland, 600-401-394, RRID:AB_2266446), Bcl-xL (Cell Signaling Technology, Cat# 2764, RRID:AB_2228008), and β-actin (Sigma-Aldrich, Cat# A5316, RRID:AB_476743) were applied to blots overnight, followed by the application of a horse-radish peroxidase-tagged secondary antibody (Bio-Rad, 1706515/1706516, Mississauga, Ontario, Canada).

Techniques:

Journal: Cell Death and Differentiation

Article Title: Mcl-1 and Bcl-xL are essential for survival of the developing nervous system

doi: 10.1038/s41418-018-0225-1

Figure Lengend Snippet: The distinct and overlapping roles of anti-apoptotic Mcl-1 and Bcl-xL in mammalian neurogenesis. Mcl-1 is required at the earlier stages of neurogenesis specifically for the survival of NPCs as they exit the cell cycle and start to differentiate. Our gene dosage data shows that Bcl-xL partially compensates for Mcl-1 in NPCs. As cells become immature neurons, Bcl-xL takes on a more prominent role, becoming the main anti-apoptotic Bcl-2 family member

Article Snippet: Primary antibodies Mcl-1 (Rockland, 600-401-394, RRID:AB_2266446), Bcl-xL (Cell Signaling Technology, Cat# 2764, RRID:AB_2228008), and β-actin (Sigma-Aldrich, Cat# A5316, RRID:AB_476743) were applied to blots overnight, followed by the application of a horse-radish peroxidase-tagged secondary antibody (Bio-Rad, 1706515/1706516, Mississauga, Ontario, Canada).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Enhanced Anti-Cancer Effects of Conditioned Medium from Hypoxic Human Adult Dermal Fibroblasts on Cervical Cancer Cells

doi: 10.3390/ijms23095134

Figure Lengend Snippet: Nodes and edges in the PPIs of intracellular proteins up- and down-regulated (> 2-fold) in HeLa cells treated with H-CM compared with N-CM.

Article Snippet: The membranes were blocked with 4% skim milk for 2 h at room temperature and then incubated with rabbit anti-human CD19 (1:3000 dilution; cat. no. CSB-RA780821A0HU; CUSABIO, Wuhan, China), CHEK1 (1:3000 dilution; cat. no. CSB-RA176809A0HU; CUSABIO), ESR1 (1:3000 dilution; cat. no. CSB-PA11399A0Rb; CUSABIO), LCK (1:3000 dilution; cat. no. CSB-PA009798; CUSABIO), MCL1 (1:3000 dilution; cat. no. CSB-PA03829A0Rb; CUSABIO), PCNA (1:3000 dilution; cat. no. CSB-PA-208009; CUSABIO), POLA1 (1:3000 dilution; cat. no. CSB-PA002170; CUSABIO), TBP (1:3000 dilution; cat. no. CSB-RA821481A0HU; CUSABIO), or TNF antibody (1:3000 dilution; cat. no. CSB-PA07427A0Rb; CUSABIO) and mouse anti-human ꞵ-actin antibody (1:10,000 dilution; cat. no. A1978; Sigma) overnight at −4 °C.

Techniques:

Journal: Oncotarget

Article Title: Targeting of BMI-1 expression by the novel small molecule PTC596 in mantle cell lymphoma

doi: 10.18632/oncotarget.25558

Figure Lengend Snippet: (A-B) BAX conformational changes, caspase-3 cleavage and Δψm loss were determined by flow cytometry in Z-138 cells (A) or REC-1 cells (B) after 18-h (BAX activation) or 20-h (caspase-3 cleavage and Δψ m loss) exposure to PTC596. The results are expressed as the mean ± SD. (C) Z-138 and REC-1 cells were treated for 24 hours with 300 nM PTC596, after which BCL-2, BCL-X L and MCL-1 protein levels were determined. In BMI-1, slow migrating phosphorylated BMI-1 bands are indicated by the open arrowhead, and non-phosphorylated BMI-1 is indicated by the closed arrowhead. The intensities of immunoblot signals were quantified and normalized to those of β-ACTIN. Levels in untreated cells were set at 1.0. Results are representative of three independent experiments.

Article Snippet: Antibodies against BMI-1, AKT, phospho-AKT (Ser473), PARP, GAPDH, and β-ACTIN were purchased from Cell Signaling Technology (Danvers, MA, USA), BCL-2 from Dako (Glostrup, Denmark), and BCL-X L and MCL-1 from BD Biosciences.

Techniques: Flow Cytometry, Activation Assay, Western Blot

Journal: Oncotarget

Article Title: Targeting of BMI-1 expression by the novel small molecule PTC596 in mantle cell lymphoma

doi: 10.18632/oncotarget.25558

Figure Lengend Snippet: (A) Z-138 cells were treated for 48 hours with 10 μM ibrutinib (IBR) and 150 nM PTC596 either as individual agents or in combination, after which BCL-2, BCL-X L and MCL-1 protein levels were determined. (B) REC-1 cells were treated with IBR (0.2 μM, 96 hours) and PTC596 (300 nM, 24 hours) either as individual agents or in combination, after which BCL-2, BCL-X L and MCL-1 protein levels were determined. PTC596 was added 72 hours after IBR exposure. (C) Z-138 cells were treated for 72 hours with IBR (2.5, 5 or 10 μM) and the selective MCL-1 inhibitor S63845 (100, 200 or 400 nM), either as individual agents or in combination, after which the annexin V-positive fractions were determined. (D) REC-1 cells were treated with IBR (0.05, 0.1 or 0.2 μM for 96 hours) and S63845 (50, 100 or 200 nM for 24 hours) either as individual agents or in combination, after which the annexin V-positive fractions were determined. The results are expressed as the mean ± SD.

Article Snippet: Antibodies against BMI-1, AKT, phospho-AKT (Ser473), PARP, GAPDH, and β-ACTIN were purchased from Cell Signaling Technology (Danvers, MA, USA), BCL-2 from Dako (Glostrup, Denmark), and BCL-X L and MCL-1 from BD Biosciences.

Techniques:

Journal: Oncotarget

Article Title: Targeting of BMI-1 expression by the novel small molecule PTC596 in mantle cell lymphoma

doi: 10.18632/oncotarget.25558

Figure Lengend Snippet: REC-1 cells were incubated for 24 hours with the indicated concentrations of ibrutinib and PTC596 either as individual agents or in combination. Viable cells were then sorted and collected into side population (SP) and non-SP cell populations, and BMI-1 and MCL-1 expression levels were determined. In BMI-1, slow migrating phosphorylated BMI-1 bands are indicated by the open arrowhead, and non-phosphorylated BMI-1 is indicated by the closed arrowhead. The intensities of immunoblot signals were quantified and normalized to those of β-ACTIN. Levels in untreated cells were set at 1.0.

Article Snippet: Antibodies against BMI-1, AKT, phospho-AKT (Ser473), PARP, GAPDH, and β-ACTIN were purchased from Cell Signaling Technology (Danvers, MA, USA), BCL-2 from Dako (Glostrup, Denmark), and BCL-X L and MCL-1 from BD Biosciences.

Techniques: Incubation, Expressing, Western Blot