anti dap3 antibody  (Santa Cruz Biotechnology)

Journal: The Journal of biological chemistry

Article Title: Oocytes orchestrate protein prenylation for mitochondrial function through selective inactivation of cholesterol biosynthesis in murine species.

doi: 10.1016/j.jbc.2023.105183

Figure Lengend Snippet: Figure 5. Excessive farnesylation of Dap3 in Ggps1-KD cells is responsible for impaired mitochondrial function. A, schematic of the screening of prenylated proteins in mitochondria of Ggps1-KD cells in vitro. Protein prenylation was assessed by immunoblotting. B and C, aqueous phase-localized and detergent phase-localized candidate proteins in mitochondrial components of control and Ggps1-KD cells were obtained via Triton X-114 extraction and analyzed by immunoblotting. Assays were performed in 293T cells (B) and Aml12 cells (C). Hsp60 served as mitochondrial markers. M: mitochondria, C: cytosol, D: detergent phase, A: aqueous phase. The experiments were repeated three times. D and E, oxygen consumption rates (OCRs) and maximal respiration in Aml12 cells were detected with an XF Cell Mito Stress Test. We cultured Aml12 cells transfected with vector, Dap3-WT, or Dap3-C363S mutant for 48 h with or without 10 μM DGBP treatment for 24 h. DGBP was used to inhibit Ggps1 activity. (n = 3, **p < 0.01 and ***p < 0.001). The experiments were repeated three times. **p < 0.01, ***p < 0.001, compared with the WT or control by Student’s t test. The data represent the mean ± SEM. See also Table S3. DGBP, digeranyl bisphosphonate; Ggps1, geranylgeranyl diphosphate synthase1.

Article Snippet: The immunoblots were blocked with 5% nonfat milk solution and incubated overnight at 4 C with primary antibodies against Ggps1 (1:400, Thermo Fisher), Rap1A (1:400, Santa Cruz), CYTB (1:1000, Proteintech), ND5 (1:1000, Proteintech), MTCO1 (1:1000, Proteintech), Tubulin (1:1000, Proteintech), β-Actin (1:1000, Proteintech), LC3 (1:1000, Proteintech), Rab7 (1:500, Santa Cruz), Rac1 (1:400, Santa Cruz), Rhoa (1:400, Santa Cruz), CLPB (1:1000, Abclonal), ABCB7 (1:1000, Abclonal), DAP3 (1:1000, Abclonal, GPD2 (1:1000, Abcam,), and HSP60 (1:1000, Proteintech).

Techniques: In Vitro, Western Blot, Control, Extraction, Cell Culture, Transfection, Plasmid Preparation, Mutagenesis, Activity Assay

Journal: International journal of oncology

Article Title: Death associated protein‑3 (DAP3) and DAP3 binding cell death enhancer‑1 (DELE1) in human colorectal cancer, and their impacts on clinical outcome and chemoresistance.

doi: 10.3892/ijo.2022.5455

Figure Lengend Snippet: Figure 1. IHC staining for the DAP3 and DELE1 in colon cancer tissues (n=52) and the adjacent tissues (n=62). (A) DAP3 expression and (B) DELE1 expres‑ sion in adjacent normal tissue, T3 and T4 colon cancer tissue with the magnification of x10 and x40, respectively. (C and D) Box plot presenting the differential (C) DAP3 and (D) DELE1 expression in adjacent tissue, early stage, and the advanced stage of colon cancer. *P<0.05, **P<0.01 and ***P<0.001. DAP3, death associated protein‑3; DELE1, DAP3 binding cell death enhancer‑1.

Article Snippet: Anti‐human DELE1 siRNA was purchased from Santa Cruz Biotechnologies, Inc. An anti‐human DAP3 ribozyme transgene that was specifically used to target and knockdown human DAP3 was prepared as previously reported (12).

Techniques: Immunohistochemistry, Expressing, Binding Assay

Journal: International journal of oncology

Article Title: Death associated protein‑3 (DAP3) and DAP3 binding cell death enhancer‑1 (DELE1) in human colorectal cancer, and their impacts on clinical outcome and chemoresistance.

doi: 10.3892/ijo.2022.5455

Figure Lengend Snippet: Figure 2. Kaplan‑Meier survival analyses showed correlations between overall survival, disease‑free survival, distant metastasis‑free survival, recurrence‑free survival and altered expression of DAP3, DELE1 and DAP3/DELE1 ratio. P‑values shown are from the log ranked test. DAP3, death associated protein‑3; DELE1, DAP3 binding cell death enhancer‑1.

Article Snippet: Anti‐human DELE1 siRNA was purchased from Santa Cruz Biotechnologies, Inc. An anti‐human DAP3 ribozyme transgene that was specifically used to target and knockdown human DAP3 was prepared as previously reported (12).

Techniques: Expressing, Binding Assay

Journal: International journal of oncology

Article Title: Death associated protein‑3 (DAP3) and DAP3 binding cell death enhancer‑1 (DELE1) in human colorectal cancer, and their impacts on clinical outcome and chemoresistance.

doi: 10.3892/ijo.2022.5455

Figure Lengend Snippet: Figure 3. Response to chemotherapy with differential transcript DELE1 and DAP3 respectively. This was based on the public database (www.rocplot.com). Shown in the figure is the median of the respective group. *P<0.05 by Mann‑Whitney U test. DAP3, death associated protein‑3; DELE1, DAP3 binding cell death enhancer‑1.

Article Snippet: Anti‐human DELE1 siRNA was purchased from Santa Cruz Biotechnologies, Inc. An anti‐human DAP3 ribozyme transgene that was specifically used to target and knockdown human DAP3 was prepared as previously reported (12).

Techniques: Mann-Whitney U-Test, Binding Assay

Journal: International journal of oncology

Article Title: Death associated protein‑3 (DAP3) and DAP3 binding cell death enhancer‑1 (DELE1) in human colorectal cancer, and their impacts on clinical outcome and chemoresistance.

doi: 10.3892/ijo.2022.5455

Figure Lengend Snippet: Figure 4. Correlation between DAP3 and DELE1 with angiogenesis biomarkers. (A and B) Correlation efficiency between DAP3 and angiogenesis markers in The Cancer Genome Atlas‑COAD cohort was presented in (A) heatmap (A) and (B) scatter plots. (C and D) Association between DELE1 and angiogenesis biomarkers was revealed in (C) heatmap and (D) scatter plots. DAP3, death associated protein‑3; DELE1, DAP3 binding cell death enhancer‑1. **P<0.01.

Article Snippet: Anti‐human DELE1 siRNA was purchased from Santa Cruz Biotechnologies, Inc. An anti‐human DAP3 ribozyme transgene that was specifically used to target and knockdown human DAP3 was prepared as previously reported (12).

Techniques: Binding Assay

Journal: International journal of oncology

Article Title: Death associated protein‑3 (DAP3) and DAP3 binding cell death enhancer‑1 (DELE1) in human colorectal cancer, and their impacts on clinical outcome and chemoresistance.

doi: 10.3892/ijo.2022.5455

Figure Lengend Snippet: Figure 5. Validation of the DAP3 and DELE1 knockdown in RKO cells by quantitative PCR gene transcript analysis. Left panel: Expression of DAP3 in the cell models. The levels of expression in wild type, control transfection (control plasmid and control siRNA), DAP3 knockdown, DELE1 knockdown and DAP3/DELE1 dual knockdown cells were compared. Control transfections resulted in minor and statistically insignificant changes of DAP3 when compared with wild type cells (P>0.05) Right panel: Expression of DELE1 in the cell models. The levels of expression in wild type, control transfection (control plasmid and control siRNA), DELE1 knockdown and DAP3/DELE1 dual knockdown cells were compared. Control transfections resulted in minor and statisti‑ cally insignificant changes of DELE1 when compared with wild type cells (P>0.05). Values presented in the figure are mean and standard deviation. Error bars represent standard deviation. **P<0.01 vs. RKO wild type cells, ΔΔP<0.001 vs. RKO‑control‑plasmid cells and ##P<0.01 VS RKO‑control‑siRNA cells. DAP3, death associated protein‑3; DELE1, DAP3 binding cell death enhancer‑1; siRNA, small interfering RNA.

Article Snippet: Anti‐human DELE1 siRNA was purchased from Santa Cruz Biotechnologies, Inc. An anti‐human DAP3 ribozyme transgene that was specifically used to target and knockdown human DAP3 was prepared as previously reported (12).

Techniques: Biomarker Discovery, Knockdown, Real-time Polymerase Chain Reaction, Expressing, Control, Transfection, Plasmid Preparation, Standard Deviation, Binding Assay, Small Interfering RNA

Journal: International journal of oncology

Article Title: Death associated protein‑3 (DAP3) and DAP3 binding cell death enhancer‑1 (DELE1) in human colorectal cancer, and their impacts on clinical outcome and chemoresistance.

doi: 10.3892/ijo.2022.5455

Figure Lengend Snippet: Figure 7. Metabolic of colorectal cancer cells following DAP3 and DELE1 knockdown. (A) Concentrations of extracellular glucose in different RKO cells were determined by luminescence measurement. (B) NAD(P)H measurements quantified by luminescence in different RKO cells. (C) Differential extracellular lactate expression and the ratio in different RKO cells. (D) NO2 Standard curve and measurements of NO2 in different RKO cells. Error bar stands standard error of means. *P<0.05 and **P<0.01 by Student's t‑test. DAP3, death associated protein‑3; DELE1, DAP3 binding cell death enhancer‑1.

Article Snippet: Anti‐human DELE1 siRNA was purchased from Santa Cruz Biotechnologies, Inc. An anti‐human DAP3 ribozyme transgene that was specifically used to target and knockdown human DAP3 was prepared as previously reported (12).

Techniques: Knockdown, Expressing, Binding Assay