anti atad2  (Cell Signaling Technology Inc)

Journal: Cellular & Molecular Biology Letters

Article Title: Effect of chronic intermittent hypoxia-induced HIF-1α/ATAD2 expression on lung cancer stemness

doi: 10.1186/s11658-022-00345-5

Figure Lengend Snippet: ATAD2 expression is upregulated in LUAD tissues and cells. A Overexpression of ATAD2 mRNA is observed in LUAD tissues compared with normal samples [gene expression profiling interactive analysis (GEPIA)]. B ATAD2 mRNA expression is significantly related to the cancer stage of the patient (GEPIA). C Prognostic feature of mRNA expression of ATAD2 in LUAD patients (Kaplan–Meier plot). Overall survival (OS) curve comparing patients with high (red) and low (black) ATAD2 expression in lung cancer plotted using the Kaplan–Meier method with a threshold of p -value < 0.05. D mRNA expression of ATAD2 is increased in lung cancer cells compared with normal lung epithelial HBE cells. E , F Protein expression of ATAD2 is increased in the tissues of lung cancer patients. * p < 0.05

Article Snippet: The primary antibodies used were anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (5174; Cell Signaling Technologies) and anti-ATAD2 (50563; Cell Signaling Technologies) antibodies.

Techniques: Expressing, Over Expression

Journal: Cellular & Molecular Biology Letters

Article Title: Effect of chronic intermittent hypoxia-induced HIF-1α/ATAD2 expression on lung cancer stemness

doi: 10.1186/s11658-022-00345-5

Figure Lengend Snippet: Effects of ATAD2 on lung cancer cell viability and proliferation. A Validation of siRNA knockdown efficiency in A549 and SPC-A1 cells as determined using RT-qPCR. B , C Cell counting kit (CCK)-8 proliferation assay in siRNA-negative control (NC) or siRNA-ATAD2-transfected A549 and SPC-A1 cells. D 5-Ethynyl-2′-deoxyuridine (EdU) proliferation assay using siRNA-NC- or siRNA-ATAD2-transfected A549 and SPC-A1 cells. Scale bar = 100 μm. E Colony-formation proliferation assay in siRNA-NC- or siRNA-ATAD2-transfected A549 and SPC-A1 cells. Error bars indicate mean ± standard deviation (SD). All experiments performed in triplicate. * p < 0.05

Article Snippet: The primary antibodies used were anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (5174; Cell Signaling Technologies) and anti-ATAD2 (50563; Cell Signaling Technologies) antibodies.

Techniques: Quantitative RT-PCR, Cell Counting, CCK-8 Assay, Proliferation Assay, Negative Control, Transfection, Standard Deviation

Journal: Cellular & Molecular Biology Letters

Article Title: Effect of chronic intermittent hypoxia-induced HIF-1α/ATAD2 expression on lung cancer stemness

doi: 10.1186/s11658-022-00345-5

Figure Lengend Snippet: Effects of ATAD2 knockdown on lung cancer cell migration, invasion, and stemness. A , B Transwell migration assay using siRNA-NC- or siRNA-ATAD2-transfected A549 and SPC-A1 cells. C , D Transwell invasion assay using siRNA-NC- or siRNA-ATAD2-transfected A549 and SPC-A1 cells. E , F Tumor sphere formation assay using siRNA-NC- or siRNA-ATAD2-transfected A549 and SPC-A1 cells. Scale bar = 100 μm. G , H CD133 and CD44 mRNA levels in siRNA-NC- or siRNA-ATAD2-transfected A549 and SPC-A1 cells. Error bars indicate mean ± SD. All experiments performed in triplicate. * p < 0.05

Article Snippet: The primary antibodies used were anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (5174; Cell Signaling Technologies) and anti-ATAD2 (50563; Cell Signaling Technologies) antibodies.

Techniques: Migration, Transwell Migration Assay, Transfection, Transwell Invasion Assay, Tube Formation Assay

Journal: Cellular & Molecular Biology Letters

Article Title: Effect of chronic intermittent hypoxia-induced HIF-1α/ATAD2 expression on lung cancer stemness

doi: 10.1186/s11658-022-00345-5

Figure Lengend Snippet: Effect of decreased ATAD2 levels on mtROS production in lung cancer cells. A ATAD2 mRNA expression in lung cancer cells transfected with the ATAD2 overexpression plasmid. B Cellular mtROS accumulation in A549 and SPC-A1 cells assessed using flow cytometry analysis. C , D Nuclei are stained with Hoechst (blue), and mitochondria ROS are stained using MitoSOX-red in A549 and SPCA1 cells. Merged panels demonstrate the number of MitoSOX-red positive cells among the total cells. Scale bar = 100 μm. Error bars indicate mean ± SD. All experiments performed in triplicate. * p < 0.05

Article Snippet: The primary antibodies used were anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (5174; Cell Signaling Technologies) and anti-ATAD2 (50563; Cell Signaling Technologies) antibodies.

Techniques: Expressing, Transfection, Over Expression, Plasmid Preparation, Flow Cytometry, Staining

Journal: Cellular & Molecular Biology Letters

Article Title: Effect of chronic intermittent hypoxia-induced HIF-1α/ATAD2 expression on lung cancer stemness

doi: 10.1186/s11658-022-00345-5

Figure Lengend Snippet: CIH-induced HIF-1α activated ATAD2 during lung cancer progression. A Gene correlation between ATAD2 and HIF-1α in LUAD determined by consulting http://timer.comp-genomics.org/ . B , C Analysis of ATAD2 promoter activity. Cells were transfected with the HIF-1α-FLAG or control plasmid for 24 h. A549 and SPC-A1 cells were transfected with an ATAD2 promoter plasmid or a plasmid containing a mutated ATAD2 promoter sequence for another 24 h. D HIF-1α, ATAD2, CD133, and CD44 mRNA expression levels in A549 cells incubated under normoxia, CIH, or CIH combined with HIF-1α. E , F Flow cytometry analysis of A549 cells transfected with siRNA-NC or siRNA-ATAD2 under normoxia or CIH. Error bars indicate mean ± SD. All experiments performed in triplicate. * p < 0.05

Article Snippet: The primary antibodies used were anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (5174; Cell Signaling Technologies) and anti-ATAD2 (50563; Cell Signaling Technologies) antibodies.

Techniques: Activity Assay, Transfection, Plasmid Preparation, Sequencing, Expressing, Incubation, Flow Cytometry