Journal: Oncotarget

Article Title: MicroRNA-145 induces apoptosis of glioma cells by targeting BNIP3 and Notch signaling

doi: 10.18632/oncotarget.18604

Figure Lengend Snippet: (A) Quantitative real-time PCR of miR-145 expression in glioma samples (n = 19) compared with normal samples (n = 10). (B) quantitative real-time PCR of miR-145 expression in rat glioma tissues (n = 6) and U87, U251 glioma cells compared to normal tissues (n = 6). (C) Quantitative real-time PCR of BNIP3 mRNA expression in glioma samples (n = 19) compared with normal samples (n = 10). (D) quantitative real-time PCR of BNIP3 mRNA expression in rat glioma tissues and U87, U251 glioma cells compared to normal tissues. (E) Pathology observation of mice brain tissues sections stained with IHC (×100, ×200, ×400). (F) Immunofluorescence with BNIP3 (green) in rat normal tissues and glioma tissues (×200).*p < 0.05, **p < 0.01 versus control group.

Article Snippet: Rabbit monoclonal antibodies against Bax, Bcl-2, Caspase-3, and p21 (Abcam, USA) were used at 1:600 dilution, rabbit monoclonal antibody against Notch 1 Hes1 (Cell Signaling Technology, USA) was used at 1:600, anti-BNIP3 antibodies (Boster, China), anti-β-actin and anti-PCNA (ZSGB-BIO, China) were diluted 1:300

Techniques: Real-time Polymerase Chain Reaction, Expressing, Staining, Immunofluorescence, Control

Journal: Oncotarget

Article Title: MicroRNA-145 induces apoptosis of glioma cells by targeting BNIP3 and Notch signaling

doi: 10.18632/oncotarget.18604

Figure Lengend Snippet: (A) Western analysis of BNIP3 and control β-actin in glioma samples compared with normal samples. (B) Western analysis of BNIP3 and control β-actin in rat glioma tissues compared to normal tissues. *p < 0.05, **p < 0.01 versus control group.

Article Snippet: Rabbit monoclonal antibodies against Bax, Bcl-2, Caspase-3, and p21 (Abcam, USA) were used at 1:600 dilution, rabbit monoclonal antibody against Notch 1 Hes1 (Cell Signaling Technology, USA) was used at 1:600, anti-BNIP3 antibodies (Boster, China), anti-β-actin and anti-PCNA (ZSGB-BIO, China) were diluted 1:300

Techniques: Western Blot, Control

Journal: Oncotarget

Article Title: MicroRNA-145 induces apoptosis of glioma cells by targeting BNIP3 and Notch signaling

doi: 10.18632/oncotarget.18604

Figure Lengend Snippet: (A) Bioinformatics analysis shows the seed sequence of miR-145 binding to the 3′-UTR of BNIP3 mRNA. (B) , (C) Quantitative real-time PCR analysis of mRNA expression of BNIP3 in U87 cells treated with miR-145 mimics and inhibitor for 48 h. (D) Western analysis of protein expression of BNIP3 in U87 and U251 cells treated with miR-145 mimics and inhibitor for 48 h. (E) Immunofluorescence with BNIP3 (green) in U87 and U251 cells after miR-145 mimics or mimics NC treatment.*p < 0.05, **p < 0.01 versus control group.

Article Snippet: Rabbit monoclonal antibodies against Bax, Bcl-2, Caspase-3, and p21 (Abcam, USA) were used at 1:600 dilution, rabbit monoclonal antibody against Notch 1 Hes1 (Cell Signaling Technology, USA) was used at 1:600, anti-BNIP3 antibodies (Boster, China), anti-β-actin and anti-PCNA (ZSGB-BIO, China) were diluted 1:300

Techniques: Sequencing, Binding Assay, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Immunofluorescence, Control

Journal: Oncotarget

Article Title: MicroRNA-145 induces apoptosis of glioma cells by targeting BNIP3 and Notch signaling

doi: 10.18632/oncotarget.18604

Figure Lengend Snippet: (A) Western analysis of BNIP3, which is localized in the nucleus or the cytoplasm in U87 and U251 cells treated with miR-145 mimics and inhibitor for 48 h. (B) Wild-type 3′-UTR of BNIP3 gene was cloned into the firefly and Renilla reporter plasmid. The BNIP3-3′UTR constructs or blank plasmid were transfected into U87 and U251 cells with control or miR-145 mimics, followed by dual luciferase assays. *p < 0.05, **p < 0.01 versus control group.

Article Snippet: Rabbit monoclonal antibodies against Bax, Bcl-2, Caspase-3, and p21 (Abcam, USA) were used at 1:600 dilution, rabbit monoclonal antibody against Notch 1 Hes1 (Cell Signaling Technology, USA) was used at 1:600, anti-BNIP3 antibodies (Boster, China), anti-β-actin and anti-PCNA (ZSGB-BIO, China) were diluted 1:300

Techniques: Western Blot, Clone Assay, Plasmid Preparation, Construct, Transfection, Control, Luciferase

Journal: Oncotarget

Article Title: MicroRNA-145 induces apoptosis of glioma cells by targeting BNIP3 and Notch signaling

doi: 10.18632/oncotarget.18604

Figure Lengend Snippet: (A, B) Transfection effect of BNIP3-siRNA or BNIP3-vector was confirmed by quantitative real-time PCR. (C) U87 cells and U251 cells were stained with Hoechst 33342 dye after BNIP3-siRNA or control treatment. (D) U87 and U251 cells were stained with Tunel after BNIP3-siRNA or control treatment. (E) U87 cell apoptosis after BNIP3-siRNA or control treatment was determined by FACS. *p < 0.05, **p < 0.01 versus control group.

Article Snippet: Rabbit monoclonal antibodies against Bax, Bcl-2, Caspase-3, and p21 (Abcam, USA) were used at 1:600 dilution, rabbit monoclonal antibody against Notch 1 Hes1 (Cell Signaling Technology, USA) was used at 1:600, anti-BNIP3 antibodies (Boster, China), anti-β-actin and anti-PCNA (ZSGB-BIO, China) were diluted 1:300

Techniques: Transfection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Staining, Control, TUNEL Assay

Journal: Oncotarget

Article Title: MicroRNA-145 induces apoptosis of glioma cells by targeting BNIP3 and Notch signaling

doi: 10.18632/oncotarget.18604

Figure Lengend Snippet: (A, B) Protein expression of Notch1, p21 and Hes1 was determined by western blot analysis in U87 and U251 cells transfected with miR-145 mimics and mimics-NC, or miR-145 inhibitor and inhibitor-NC. (C, D) Western analysis of Notch1-related proteins in U87 and U251 cells transfected with Bnip3 siRNA and siRNA control, or BNIP3 expression vector and blank vector. *p < 0.05, **p < 0.01 versus control group.

Article Snippet: Rabbit monoclonal antibodies against Bax, Bcl-2, Caspase-3, and p21 (Abcam, USA) were used at 1:600 dilution, rabbit monoclonal antibody against Notch 1 Hes1 (Cell Signaling Technology, USA) was used at 1:600, anti-BNIP3 antibodies (Boster, China), anti-β-actin and anti-PCNA (ZSGB-BIO, China) were diluted 1:300

Techniques: Expressing, Western Blot, Transfection, Control, Plasmid Preparation

Journal: Oncotarget

Article Title: MicroRNA-145 induces apoptosis of glioma cells by targeting BNIP3 and Notch signaling

doi: 10.18632/oncotarget.18604

Figure Lengend Snippet: (A) Protein expression of Notch1, p21, Hes1 was determined by western blot analysis in U87 and U251 cells co-transfected with miR-145 inhibitor and BNIP3-siRNA, or with miR-145 inhibitor. *p < 0.05, **p < 0.01 versus control group.

Article Snippet: Rabbit monoclonal antibodies against Bax, Bcl-2, Caspase-3, and p21 (Abcam, USA) were used at 1:600 dilution, rabbit monoclonal antibody against Notch 1 Hes1 (Cell Signaling Technology, USA) was used at 1:600, anti-BNIP3 antibodies (Boster, China), anti-β-actin and anti-PCNA (ZSGB-BIO, China) were diluted 1:300

Techniques: Expressing, Western Blot, Transfection, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: miR-103 Regulates Oxidative Stress by Targeting the BCL2/Adenovirus E1B 19 kDa Interacting Protein 3 in HUVECs

doi: 10.1155/2015/489647

Figure Lengend Snippet: Validation of BNIP3 as a miR-103 target gene. (a) The 3′-UTR of BNIP3 and mutant 3′-UTR sequences that abolished binding. (b) Luciferase activity was assessed in HUVECs transfected with BNIP3 3′-UTR-WT or BNIP3 3′-UTR-MUT and the mimic control or miR-103. (c) Luciferase activity was assessed in HUVECs transfected with BNIP3 3′-UTR-WT or BNIP3 3′-UTR-MUT and the inhibitor control or miR-103 inhibitor. (d) BNIP3 mRNA levels analyzed by qRT-PCR. (e) Western blot analysis of the endogenous expression of BNIP3 upon forced expression of miR-103. (f) The protein expression of BNIP3 in HUVECs transfected with the miR-103 inhibitor or inhibitor control was determined by western blotting. ∗ p < 0.05; ∗∗ p < 0.01.

Article Snippet: Fifty micrograms of total proteins were electrophoresed by SDS-PAGE and the proteins were transferred onto 0.22 μ m polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, MA, USA), followed by blocking with 8% nonfat milk at room temperature for 1 h. The membranes were incubated overnight at 4°C with a rabbit polyclonal anti-BNIP3 antibody (Abnova, Taipei, Taiwan) or anti-GAPDH antibody (Cwbiotech, Shanghai, China), followed by the corresponding horseradish peroxidase- (HRP-) conjugated secondary antibodies (Cwbiotech).

Techniques: Biomarker Discovery, Mutagenesis, Binding Assay, Luciferase, Activity Assay, Transfection, Control, Quantitative RT-PCR, Western Blot, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: miR-103 Regulates Oxidative Stress by Targeting the BCL2/Adenovirus E1B 19 kDa Interacting Protein 3 in HUVECs

doi: 10.1155/2015/489647

Figure Lengend Snippet: H 2 O 2 upregulated the expression of BNIP3 in HUVECs. (a) BNIP3 protein expression in HUVECs cultured in 200 μ M H 2 O 2 for different time periods (left). Quantification of BNIP3 by band densitometry analysis (right). (b) BNIP3 protein expression in HUVECs cultured with H 2 O 2 at the indicated concentration for 6 h (left). Quantification of BNIP3 by band densitometry analysis (right). (c) Western blot analysis of BNIP3 protein expression in HUVECs pretreated with or without salidroside and induced by H 2 O 2 . (d) Western blot analysis of BNIP3 protein expression in HUVECs stably expressing miR-103 treated with H 2 O 2 . GAPDH was used as the loading control. ∗ p < 0.05; ∗∗ p < 0.01.

Article Snippet: Fifty micrograms of total proteins were electrophoresed by SDS-PAGE and the proteins were transferred onto 0.22 μ m polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, MA, USA), followed by blocking with 8% nonfat milk at room temperature for 1 h. The membranes were incubated overnight at 4°C with a rabbit polyclonal anti-BNIP3 antibody (Abnova, Taipei, Taiwan) or anti-GAPDH antibody (Cwbiotech, Shanghai, China), followed by the corresponding horseradish peroxidase- (HRP-) conjugated secondary antibodies (Cwbiotech).

Techniques: Expressing, Cell Culture, Concentration Assay, Western Blot, Stable Transfection, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: miR-103 Regulates Oxidative Stress by Targeting the BCL2/Adenovirus E1B 19 kDa Interacting Protein 3 in HUVECs

doi: 10.1155/2015/489647

Figure Lengend Snippet: Cell viability and DCF analysis of HUVECs exposed to H 2 O 2 with or without BNIP3 knockdown. (a) Western blot analysis showing levels of BNIP3 when HUVECs were transfected with different BNIP3 siRNA constructs or a negative control (NC) siRNA. (b) The CCK-8 assay was performed to evaluate the cell viability of HUVECs treated as indicated. (c) An apoptosis assay was used to assess the apoptosis levels of HUVECs treated as indicated. (d) Intracellular formation of ROS was measured in HUVECs treated as indicated. (e) BNIP3 expression was determined in HUVECs transfected with siRNA and treated with or without H 2 O 2 . ∗ p < 0.05, ∗∗ p < 0.01 versus control and # p < 0.05 versus H 2 O 2 group.

Article Snippet: Fifty micrograms of total proteins were electrophoresed by SDS-PAGE and the proteins were transferred onto 0.22 μ m polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, MA, USA), followed by blocking with 8% nonfat milk at room temperature for 1 h. The membranes were incubated overnight at 4°C with a rabbit polyclonal anti-BNIP3 antibody (Abnova, Taipei, Taiwan) or anti-GAPDH antibody (Cwbiotech, Shanghai, China), followed by the corresponding horseradish peroxidase- (HRP-) conjugated secondary antibodies (Cwbiotech).

Techniques: Knockdown, Western Blot, Transfection, Construct, Negative Control, CCK-8 Assay, Apoptosis Assay, Expressing, Control