Journal: The Journal of Cell Biology

Article Title: PINK1 drives Parkin self-association and HECT-like E3 activity upstream of mitochondrial binding

doi: 10.1083/jcb.201210111

Figure Lengend Snippet: C431S Parkin activity in cells. (A) HeLa cells expressing either WT, C431S, or C431F YFP–Parkin were treated with 10 µM CCCP for 3 h. Cells were immunostained using antibodies against the mitochondrial marker HtrA2/Omi. The graph represents means ± SD of counts in >150 cells per sample in three independent experiments of YFP–Parkin colocalizing with mitochondria. Bars, 10 µm. (B) HeLa cells were cotransfected with untagged Parkin variants as in A and EGFP-N1 in a 3:1 ratio. Cells were treated with 10 µM CCCP for 24 h and immunostained for the matrix protein Trap1. The graph represents scoring of cells positive for GFP and negative for mitochondria by lack of Trap1 signal. More than 150 cells were counted per sample in three replicates examined on at least two separate occasions. Error bars represent means ± SD. Bars, 10 µm. (C) HeLa cells were transfected with or without PINK1 siRNA for 24 h before transfection with untagged C431F or C431S Parkin for 24 h followed by treatment with DMSO or 10 µM CCCP for 3 h. Cytosol fractions were heat denatured with 1% SDS at 90°C and then split and treated with or without 100 mM NaOH at 37°C for 1 h. Samples were analyzed by immunoblotting using Parkin, PINK1, and β-actin antibodies. *, nonspecific band.

Article Snippet: Antibodies used in this study were Mfn1 (rabbit polyclonal antibodies made in house), HtrA2/Omi (R&D Systems), VDAC1 (EMD Millipore), MitoNEET (Proteintech), Parkin (Santa Cruz Biotechnology, Inc.), TRAP1 (Abcam), PINK1 (Novus Biologicals), Fis1 (Enxo Life Sciences), Tom70 (Abcam), Bak (Abcam), K48 and K63 linkage specific (EMD Millipore), polyubiquitin FK1 (Enzo Life Sciences), and Mfn2 (a kind gift from Mike Ryan, La Trobe University, Melbourne, Australia).

Techniques: Activity Assay, Expressing, Marker, Transfection, Western Blot

Journal: Cancer letters

Article Title: Restricting metabolic plasticity enhances stress adaptation through the modulation of PDH and HIF1A in TRAP1-depleted colon cancer

doi: 10.1016/j.canlet.2025.217977

Figure Lengend Snippet: (A)TRAP1 expression levels in normal versus colon tumor tissues, according to the TCGA database, as displayed on the GEP2 website. (B) TRAP1 protein expression was not detectable in TRAP1 CRISPR/Cas9 CT26 cells using Western blot analysis. (C) TRAP1 mRNA levels were quantified by qPCR in WT and KO colon cancer cells. (D) Cell growth of WT and KO cells was measured at 24 h, 48 h, 72 h using CCK-8. (E) GSEA of proteomic data using Molecular Signatures Database (MSigDB) GO BP gene set is summarized as the normalized enrichment score (NES) in WT and KO cells. (F) GSEA plots of cytoplasm translation, regulation of cell cycle G1-S, detoxification and cellular oxidant detoxification gene signatures that are associated with depletion of TRAP1. (G) Intracellular ROS levels in WT and KO cells were detected using the abcam cellular ROS assay kit and analyzed by flow cytometry. (H) Quantitation of ROS. (I) Cell lysates were prepared from WT EV, KO2 EV and KO2 TOE for Western blot analysis to detect TRAP1 and Actin proteins. (J) Intracellular ROS levels in WT EV, KO2 EV and KO2 TOE were detected using an ROS assay kit and analyzed by flow cytometry. (K) Quantitation of ROS. P < 0.05 (*), P < 0.01 (**). (L) WT and individual KO cells were collected and stained with PI. The cell cycle was analyzed using flow cytometry and BD software. (M) Cells were seeded in 6-well plates and cultured for 7 days. Cells were stained with crystal violet, and colony numbers were quantified in (N).

Article Snippet: TRAP1 CRISPR/Cas9 plasmid was obtained from Santa Cruz (Dallas, TX, USA). pCMV6 and pCMV6/TRAP1 plasmids were obtained from Origene (Rockville, MD, USA).

Techniques: Expressing, CRISPR, Western Blot, CCK-8 Assay, ROS Assay, Flow Cytometry, Quantitation Assay, Staining, Software, Cell Culture

Journal: Cancer letters

Article Title: Restricting metabolic plasticity enhances stress adaptation through the modulation of PDH and HIF1A in TRAP1-depleted colon cancer

doi: 10.1016/j.canlet.2025.217977

Figure Lengend Snippet: (A, B) The pH value of the culture medium was measured in WT and KO CT26 cells. (C, D) Enrichment analysis of proteomic data was performed using the GSEA MSigDB GO BP gene set. GSEA plots and heat maps for pH regulation genes in the TRAP1 metabolic gene signature. (E) GSEA of mass spectrometry was performed with MSigDB GO CC gene set and is summarized as the normalized enrichment score (NES) in WT and KO cells. (F) GSEA plots for NADH dehydrogenase complex and cytosolic ribosome are presented within the TRAP1 metabolic gene signature. (G, H) The oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) of WT and KO cells were measured using the Seahorse assay. The data were analyzed using Wave software. (I) The energy map generated from the Seahorse assay in WT and KO cells is shown. The empty and dotted squares represent duplicate experiments within each group. Blue indicates the WT group, red represents the KO6 group, and green represents the KO7 group. (J, K) Glucose consumption and lactate production were measured in WT and KO cells using Glucose-GLO and Lactate-GLO kits, respectively. (L) Illustration of TRAP1 depletion favors the glycolysis pathway than oxidative phosphorylation in colon cancer. P < 0.05 (*), P < 0.01 (**).

Article Snippet: TRAP1 CRISPR/Cas9 plasmid was obtained from Santa Cruz (Dallas, TX, USA). pCMV6 and pCMV6/TRAP1 plasmids were obtained from Origene (Rockville, MD, USA).

Techniques: Mass Spectrometry, Software, Generated, Phospho-proteomics

Journal: Cancer letters

Article Title: Restricting metabolic plasticity enhances stress adaptation through the modulation of PDH and HIF1A in TRAP1-depleted colon cancer

doi: 10.1016/j.canlet.2025.217977

Figure Lengend Snippet: (A) TRAP1, PDK1, pPDH (S232), pPDH (S293), PDH, and Actin protein levels were detected in WT and individual KO cells using Western blot analysis. (B) WT and individual KO cells were fixed with 4 % paraformaldehyde and stained with pPDH (S232) antibody. Staining was analyzed using a Nikon microscope. (C) WT and KO cells were treated with various concentrations of rotenone for 48 h. Cell viability was determined using the CCK-8 assay at OD450. (D) Cells were treated with either Ctrl or 200 nM rotenone for 24 h. Intracellular ROS levels were detected using the abcam cellular ROS assay kit and analyzed by flow cytometry. Quantitation of intracellular ROS levels is shown in (E). (F, G) Glucose consumption and lactate production were measured in cells treated with 10 nM or 50 nM rotenone using Glucose-GLO and Lactate-GLO kits, respectively. (H) WT and KO cells were treated with either Ctrl, 1.25 mM, or 2.5 mM 2DG for 7 days. Cells were stained with crystal violet, and colony numbers were quantified in (I). (J) WT and KO cells were treated with various concentrations of 2DG for 48 h. Cell viability was determined using the CCK-8 assay at OD450. (K) WT and KO cells were treated with Ctrl or 5 mM 2DG for 24 h. Cellular ROS levels were detected using the abcam cellular ROS assay kit and analyzed by flow cytometry. Quantitation of cellular ROS levels is shown in (L). (M) WT and KO cells were treated with 0.625 mM, 1.25 mM, or 2.5 mM 2DG for 24 h. The culture medium was collected, and glucose consumption was analyzed using Promega Glucose-GLO kits. (N) WT and KO cells were treated with various concentrations of 2DG for 48 h. Cell lysates were prepared for Western blot analysis to detect TRAP1, CHOP, PARP, and Actin protein levels. P < 0.05 (*), P < 0.01 (**).

Article Snippet: TRAP1 CRISPR/Cas9 plasmid was obtained from Santa Cruz (Dallas, TX, USA). pCMV6 and pCMV6/TRAP1 plasmids were obtained from Origene (Rockville, MD, USA).

Techniques: Western Blot, Staining, Microscopy, CCK-8 Assay, ROS Assay, Flow Cytometry, Quantitation Assay

Journal: Cancer letters

Article Title: Restricting metabolic plasticity enhances stress adaptation through the modulation of PDH and HIF1A in TRAP1-depleted colon cancer

doi: 10.1016/j.canlet.2025.217977

Figure Lengend Snippet: (A) GSEA of mass spectrometry was performed with MSigDB HALLMARK gene set and is summarized as the normalized enrichment score (NES) in WT and KO cells. (B, C) Enrichment analysis was carried out using the GSEA HALLMARK gene set. GSEA plots of hypoxia and glycolysis genes are presented within the TRAP1 metabolic gene signature. (D) Differential gene expressions were analyzed using DEseq2, with fold change >1.5 and FDR <0.1. Volcano plot shows relative fold change (log2) in protein abundance versus −log10(P values) from WT cells compared with KO cells. Proteins that demonstrate a significant change in expression are colored, with decreased expression in green color and increased expression in red color. (E, F) BP and KEGG pathway analysis of the differential gene expressions was conducted using ShinyGO website. The dot plot represents the top 10 significant pathways ranked according to − log enrichment P value. (G) Correlation analysis of TRAP1 with HIF1A and HIF1A with MCT1 expression in the TCGA database was performed using the ENCORI website. (H) Cells were fixed with 4 % paraformaldehyde and stained with HIF1A antibody. Staining was analyzed ushing a Nikon microscope. (I) Nuclear and cytoplasmic fractions of WT and KO cells were separated using a nuclear extraction kit (ThermoFisher). TRAP1, HIF1A, Lamin B1, and Tubulin proteins were detected using specific antibodies by Western blotting. (J) WT and KO cells stably expressing HRE-luciferase were subjected to a reporter assay using the Promega luciferase kit, One-GLO. Luminescence levels were measured using a Luminescence reader. (K) TRAP1, HIF1A, GLUT1, MCT1, and Actin proteins were detected using Western blotting. (L) Illustration of TRAP1 depletion induces ROS generation to facilitate glycolysis pathway through ROS-HIF1A axis. P < 0.05 (*), P < 0.01 (**).

Article Snippet: TRAP1 CRISPR/Cas9 plasmid was obtained from Santa Cruz (Dallas, TX, USA). pCMV6 and pCMV6/TRAP1 plasmids were obtained from Origene (Rockville, MD, USA).

Techniques: Mass Spectrometry, Quantitative Proteomics, Expressing, Staining, Microscopy, Extraction, Western Blot, Stable Transfection, Luciferase, Reporter Assay

Journal: Cancer letters

Article Title: Restricting metabolic plasticity enhances stress adaptation through the modulation of PDH and HIF1A in TRAP1-depleted colon cancer

doi: 10.1016/j.canlet.2025.217977

Figure Lengend Snippet: (A) Cell lysates were prepared from WT EV, KO2 EV, KO2 TOE, KO6 EV, and KO6 TOE cells. TRAP1, PDK1, pPDH (S232), pPDH (S293), PDH, and Actin proteins were detected using Western blot analysis. (B) Cells were fixed with 4 % paraformaldehyde and stained with pPDH (S232) antibody. Staining was analyzed using a Nikon microscope. (C) Cells were treated with different dosages of rotenone for 48 h. Cell viability was analyzed using the CCK-8 assay at OD450. (D) WT EV, KO2 EV and KO2 TOE cells were treated with Ctrl and 200 nM rotenone for 24 h. Intracellular ROS levels of were detected using the abcam cellular ROS assay kit and analyzed by flow cytometry. (E) Quantitation of intracellular ROS levels. (F) Cells were fixed with 4 % paraformaldehyde and stained with HIF1A antibody. Staining was analyzed using a Nikon microscope. (G) Cell lysates were prepared from WT EV, KO2 EV and KO2 TOE cells. TRAP1, GLUT1, MCT1, and Actin protein levels were detected using western blot analysis. (H, I) The relative glucose consumption rate and lactate concentration of the culture medium at different time points in WT EV, KO2 EV and KO2 TOE cells were measured using Glucose-GLO and Lactate-GLO kits. (J) Cells were treated with 5 mM 2DG for 24 h. Intracellular ROS levels were detected using the abcam cellular ROS assay kit and analyzed by flow cytometry. (K) Quantitation of intracellular ROS levels. (L) Cells were treated with different dosages of 2DG for 48 h. Cell viability was analyzed using the CCK-8 assay at OD450. P < 0.05 (*), P < 0.01 (**).

Article Snippet: TRAP1 CRISPR/Cas9 plasmid was obtained from Santa Cruz (Dallas, TX, USA). pCMV6 and pCMV6/TRAP1 plasmids were obtained from Origene (Rockville, MD, USA).

Techniques: Western Blot, Staining, Microscopy, CCK-8 Assay, ROS Assay, Flow Cytometry, Quantitation Assay, Concentration Assay

Journal: Cancer letters

Article Title: Restricting metabolic plasticity enhances stress adaptation through the modulation of PDH and HIF1A in TRAP1-depleted colon cancer

doi: 10.1016/j.canlet.2025.217977

Figure Lengend Snippet: (A) WT and KO cells were treated with 2 mM Trolox for 2 h. Intracellular ROS levels were detected using the abcam cellular ROS assay kit and analyzed by flow cytometry. (B) Quantitation of intracellular ROS levels were performed. (C) Cells were treated with Ctrl and 1 mM Trolox for 24h. Cells were fixed with 4 % paraformaldehyde and stained with HIF1A antibody. Staining was analyzed using a Nikon microscope. (D) WT and KO cells stably expressing HRE-luciferase were treated with various concentrations of Trolox for 18 h. Reporter assays were performed using One-GLO. (E) Cells were treated with 0.3 mM or 1 mM Trolox for 24 h. Cell lysates were prepared for Western blot analysis to detect TRAP1, HIF1A, GLUT1, MCT1, and Actin proteins. (F) Cells were treated with 0.3 mM or 1 mM Trolox for 48 h. The pH value of the culture medium was measured using a pH meter. (G, H) Cells were treated with Ctrl, 2 mM Trolox for 24 h. Glucose consumption and lactate production in Trolox-treated cells were measured using Glucose-GLO and Lactate-GLO kits. (I) Cells were treated with Ctrl, 50 μM, or 100 μM PX478 for 24 h. Cell lysates were prepared for Western blot analysis to detect TRAP1, HIF1A, GLUT1, MCT1, and Actin proteins. (J, K) Glucose consumption and lactate production in 100 μM PX-478-treated cells were measured using Glucose-GLO and Lactate-GLO kits. (L) Cells were treated with 100 μM PX478 for 24 h. Cells were fixed with 4 % paraformaldehyde and stained with pPDH(S232) antibody. Staining was analyzed using a Nikon microscope. (M) Cells were treated with 100 μM PX478 for 48 h. Cell lysates were prepared for Western blot analysis to detect TRAP1, HIF1A, PDK1, pPDH(S232), PDH, and Actin proteins. P < 0.05 (*), P < 0.01 (**).

Article Snippet: TRAP1 CRISPR/Cas9 plasmid was obtained from Santa Cruz (Dallas, TX, USA). pCMV6 and pCMV6/TRAP1 plasmids were obtained from Origene (Rockville, MD, USA).

Techniques: ROS Assay, Flow Cytometry, Quantitation Assay, Staining, Microscopy, Stable Transfection, Expressing, Luciferase, Western Blot

Journal: Cancer letters

Article Title: Restricting metabolic plasticity enhances stress adaptation through the modulation of PDH and HIF1A in TRAP1-depleted colon cancer

doi: 10.1016/j.canlet.2025.217977

Figure Lengend Snippet: (A) Cells were treated with 20 mM DCA for 48 h. TRAP1, PDK1, pPDH (S232), pPDH (S293), PDH, and Actin protein levels were detected using Western blot analysis. (B) Cells were treated with Ctrl and 20 mM DCA for 24 h. Cells were fixed with 4 % paraformaldehyde and stained with pPDH (S232) antibody. Staining was analyzed using a Nikon microscope. (C, D) The pH value of the culture medium was measured in cells treated with various concentrations of DCA. (E, F) The relative glucose consumption rate and lactic acid concentration of the culture medium in DCA-treated cells were measured using Glucose-GLO and Lactate-GLO kits. (G) WT and KO cells were treated with Ctrl, 10 mM DCA, or 20 mM DCA for 24 h. Intracellular ROS levels were detected using the abcam cellular ROS assay kit and analyzed by flow cytometry. (H) Quantitation of intracellular ROS levels. (I) Cells were treated with Ctrl, 10 mM, 20 mM, or 40 mM DCA for 48 h. Cell viability was analyzed using the CCK-8 assay at OD450. (J) Cells were treated with Ctrl, 10 mM, or 20 mM DCA for 7 days. Cells were stained with crystal violet. Colony numbers were quantified in (K). (L) Illustration of the DCA activated the PDH activity to suppress cell viability in KO cells. P < 0.05 (*), P < 0.01 (**).

Article Snippet: TRAP1 CRISPR/Cas9 plasmid was obtained from Santa Cruz (Dallas, TX, USA). pCMV6 and pCMV6/TRAP1 plasmids were obtained from Origene (Rockville, MD, USA).

Techniques: Western Blot, Staining, Microscopy, Concentration Assay, ROS Assay, Flow Cytometry, Quantitation Assay, CCK-8 Assay, Activity Assay

Journal: Oxidative medicine and cellular longevity

Article Title: Tumor Necrosis Factor Receptor-Associated Protein 1 Protects against Mitochondrial Injury by Preventing High Glucose-Induced mPTP Opening in Diabetes.

doi: 10.1155/2020/6431517

Figure Lengend Snippet: Figure 3: Effects of TRAP1 on mitochondrial morphology and dysfunction in NRK-52e cells after high-glucose injury. (a) Mitochondrial morphology was observed by TEM. Arrows indicate mitochondria; scale bar, 500 nm. (b) ATP depletion was measured using firefly luciferase. (c, d) Representative plots of MMP and statistical analyses were determined by flow cytometric analysis of tetramethylrhodamine ethyl ester-labeled NRK-52e cells. (e, f) Representative plots and statistical analysis of intracellular ROS in cells labeled with the fluorescent probe CellROX Deep Red and analyzed by flow cytometry. (g, h) Typical fluorescence photomicrograph and quantitative analysis of mitochondrial superoxide (red: MitoSox; blue: DAPI); scale bar, 50 μm. ROS: reactive oxygen species. The results are presented as the mean ± SEM; n = 3, ∗p < 0:05, ∗∗p < 0:01, and ∗∗∗p < 0:001 for each pair of groups indicated.

Article Snippet: After being blocked in 5% BSA for 1 h, the membranes were probed overnight at 4°C with antibodies against TRAP1 (1 : 1000; Novus Biologicals, USA) and GAPDH (1 : 5000; Proteintech, USA).

Techniques: Luciferase, Labeling, Cytometry

Journal: Oxidative medicine and cellular longevity

Article Title: Tumor Necrosis Factor Receptor-Associated Protein 1 Protects against Mitochondrial Injury by Preventing High Glucose-Induced mPTP Opening in Diabetes.

doi: 10.1155/2020/6431517

Figure Lengend Snippet: Figure 4: TRAP1 protects against high-glucose-induced mitochondrial dysfunction via mPTP opening regulation. (a, b) Representative graphs and statistical analyses of mPTP by flow cytometry with the fluorescent probe calcein AM labeling. (c) Mitochondrial ultrastructural damage was observed by TEM. Arrows indicate mitochondria; scale bar, 500 nm. (d, e) Typical graphs of mitochondrial membrane potential and statistical analysis determined with tetramethylrhodamine ethyl ester labeling by flow cytometric analysis at 48 h. (f, g) Representative graphs and statistical analysis of mPTP by flow cytometry. mPTP: mitochondrial permeability transition pore. The results are presented as the mean ± SEM; n = 3, ∗p < 0:05, and ∗∗p < 0:01 for each pair of groups indicated.

Article Snippet: After being blocked in 5% BSA for 1 h, the membranes were probed overnight at 4°C with antibodies against TRAP1 (1 : 1000; Novus Biologicals, USA) and GAPDH (1 : 5000; Proteintech, USA).

Techniques: Cytometry, Labeling, Membrane, Permeability

Journal: Oxidative medicine and cellular longevity

Article Title: Tumor Necrosis Factor Receptor-Associated Protein 1 Protects against Mitochondrial Injury by Preventing High Glucose-Induced mPTP Opening in Diabetes.

doi: 10.1155/2020/6431517

Figure Lengend Snippet: Figure 6: Effect of TRAP1 overexpression on biochemical parameters and histopathology in vivo. Rats were treated with an injection of STZ to induce diabetes and AAV 2/9 to overexpress TRAP1. Rats transfected with empty-GFP vectors (AAV-VE) are shown for comparison. Blood and kidney tissues were collected after 12 weeks. (a) Western blot graphs and densitometric analyses of TRAP1 expression in kidneys. GAPDH was used as a control. (b–f) Blood glucose (b), serum FRU (c), serum Ccr (d), urine albumin to creatinine (e), and serum urea (f) levels were analyzed. (g) Representative histology of the renal cortex and outer medulla: H&E staining, PAS for glycogen, Sirius red staining for the detection of fibrosis, and Masson’s trichrome staining for connective tissue; scale bar: 50 μm. The results are presented as the mean ± SEM; n = 4‐6, ∗p < 0:05, ∗∗p < 0:01, and ∗∗∗p < 0:001 for each pair of groups indicated.

Article Snippet: After being blocked in 5% BSA for 1 h, the membranes were probed overnight at 4°C with antibodies against TRAP1 (1 : 1000; Novus Biologicals, USA) and GAPDH (1 : 5000; Proteintech, USA).

Techniques: Over Expression, Histopathology, In Vivo, Injection, Transfection, Comparison, Western Blot, Expressing, Control, Staining

Journal: Oxidative medicine and cellular longevity

Article Title: Tumor Necrosis Factor Receptor-Associated Protein 1 Protects against Mitochondrial Injury by Preventing High Glucose-Induced mPTP Opening in Diabetes.

doi: 10.1155/2020/6431517

Figure Lengend Snippet: Figure 7: Effects of TRAP1 on diabetes-induced apoptosis and mitochondrial damage in rat kidney. (a) Representative images of (TUNEL) staining (red: TUNEL; blue: DAPI); scale bar, 100 μm. (b) Quantification of TUNEL-positive cells in fields. (c) Mitochondria of tubular epithelial cells viewed by TEM; scale bar, 1 μm. (d) ATP levels measured by firefly luciferase detection. (e, f) Representative photomicrographs and quantification of ROS staining in rat renal tubules; scale bar, 100 μm. The results are presented as the mean ± SEM; n = 4‐6, ∗p < 0:05, ∗∗p < 0:01, and ∗∗∗p < 0:001 for each pair of groups indicated.

Article Snippet: After being blocked in 5% BSA for 1 h, the membranes were probed overnight at 4°C with antibodies against TRAP1 (1 : 1000; Novus Biologicals, USA) and GAPDH (1 : 5000; Proteintech, USA).

Techniques: TUNEL Assay, Staining, Luciferase

Journal: Oncology Letters

Article Title: Luteolin potentiates low-dose oxaliplatin-induced inhibitory effects on cell proliferation in gastric cancer by inducing G 2 /M cell cycle arrest and apoptosis

doi: 10.3892/ol.2021.13134

Figure Lengend Snippet: Changes in TRAP1, P-ERK1/2/ERK1/2 and CDC25C protein expression levels in mouse forestomach carcinoma cells induced by luteolin (20 µM) and/or oxaliplatin (5 µM). (A-C) TRAP1, P-ERK1/2/ERK1/2, CDC25C and β-actin protein expression levels were assessed by western blot analysis, and quantitative analysis of protein expression levels is shown in the histogram. *P<0.05, **P<0.01. Experiments were repeated at least in triplicate. Lut, luteolin; Oxa, oxaliplatin; TRAP1, tumor necrosis factor receptor-associated protein 1; ERK1/2, extracellular-regulated protein kinases1/2; CDC25C, cell division cycle 25 homolog C.

Article Snippet: The primary antibodies used were: β-actin mouse monoclonal antibody (cat. no. TA-09; 1:2,000; OriGene Technologies, Inc.), Bcl-2 rabbit polyclonal antibody (cat. no. ab196495, 1:1,000; Abcam), BCL-2-associated X protein (Bax) rabbit monoclonal antibody (cat. no. ab182734; 1:1,000; Abcam), cyclin A2 rabbit monoclonal antibody (cat. no. ab181591; 1:2,000; Abcam), cyclin B1 rabbit monoclonal antibody (cat. no. ab32053; 1:1,000; Abcam), cyclin-dependent kinase-1 (CDK1) rabbit monoclonal antibody (cat. no. ab133327; 1:20,000; Abcam), tumor necrosis factor receptor-associated protein 1 (TRAP1) rabbit polyclonal antibody (cat. no. 10325-1-AP; 1:2,000; ProteinTech Group, Inc.), cell division cycle 25 homolog C (CDC25C) mouse monoclonal antibody (cat. no. 66912-1-lg; 1:2,000; ProteinTech Group, Inc.), extracellular-regulated protein kinases1/2 (ERK1/2) rabbit polyclonal antibody (cat. no. 9102s; 1:1,000; Cell Signaling Technology, Inc.) and p-ERK1/2 mouse monoclonal antibody (cat. no. 9106s; 1:1,000; Cell Signaling Technology, Inc.).

Techniques: Expressing, Western Blot

Journal: eLife

Article Title: Intrinsic OXPHOS limitations underlie cellular bioenergetics in leukemia

doi: 10.7554/eLife.63104

Figure Lengend Snippet:

Article Snippet: Transfected construct ( Homo sapien ) , shRNA to TRAP1 , Origene , CAT#: TL300868V , shRNA lentiviral particles packaged from pGFP-C-shLenti vector.

Techniques: Isolation, Lysis, Transfection, Construct, shRNA, Plasmid Preparation, Sequencing, Recombinant, Software