Journal: American Journal of Translational Research

Article Title: Knockdown of miR-27a reduces TGFβ-induced EMT and H 2 O 2 -induced oxidative stress through regulating mitochondrial autophagy

doi:

Figure Lengend Snippet: miR-27a mimic decreases cell proliferation and increases apoptosis in ESCs. Mimic-NC, mimic-27, inhibitor-NC, and inhibitor-27 were transfected into ESCs, respectively. A. RNA level of mir-27a was examined by qPCR. B. Cell proliferation was examined by CCK8 assay. C. Flow cytometry was carried out for cell apoptosis examination. Data were presented as mean ± SD. **P<0.01, ***P<0.001.

Article Snippet: Flow cytometric analysis Following the manufacturer’s protocol, we determined cell apoptosis by flow cytometric analysis using an Annexin-V-FITC Apoptosis Detection kit (Solarbio, China).

Techniques: Transfection, CCK-8 Assay, Flow Cytometry

Journal: American Journal of Translational Research

Article Title: Knockdown of miR-27a reduces TGFβ-induced EMT and H 2 O 2 -induced oxidative stress through regulating mitochondrial autophagy

doi:

Figure Lengend Snippet: Downregulating miR-27a reduced H2O2-induced oxidative stress. A. ESCs were treated with or without H2O2. The miR-27a RNA level was examined by qPCR. ESCs, transfected with or without inhibitor-NC and inhibitor-27, were then intervened by 100 μM H2O2. B. Intracellular ROS production was detected with DCFH-DA assay. C. Western blotting was conducted to examine Bcl-2 and Bax protein levels. D. Flow cytometry was carried out for cell apoptosis examination. E. Total p65 and phosphorylated p65 levels was accessed by western blotting. Data were presented as mean ± SD. **P<0.01, ***P<0.001.

Article Snippet: Flow cytometric analysis Following the manufacturer’s protocol, we determined cell apoptosis by flow cytometric analysis using an Annexin-V-FITC Apoptosis Detection kit (Solarbio, China).

Techniques: Transfection, DCFH-DA Assay, Western Blot, Flow Cytometry

Journal: American Journal of Translational Research

Article Title: Knockdown of miR-27a reduces TGFβ-induced EMT and H 2 O 2 -induced oxidative stress through regulating mitochondrial autophagy

doi:

Figure Lengend Snippet: miR-27a regulates mitophagy in H2O2-treated ESCs. A. Mimic-NC, mimic-27, inhibitor-NC, and inhibitor-27 were transfected into ESCs, respectively. PINK1, Parkin, and LC3 protein levels were examined by western blotting. ESCs, transfected with inhibitor-NC, inhibitor-27, and inhibitor-27+si-PINK1, were then intervened by 100 μM H2O2. B. PINK1, Parkin, and LC3 protein levels were examined by western blotting. C. Intracellular ROS production was detected by DCFH-DA assay. D. Western blotting was carried out to determine Bcl-2 and Bax protein levels. E. Cell apoptosis was examined by flow cytometry. Data were presented as mean ± SD. *, **, and *** represent P<0.05, P<0.01, and P<0.001, respectively.

Article Snippet: Flow cytometric analysis Following the manufacturer’s protocol, we determined cell apoptosis by flow cytometric analysis using an Annexin-V-FITC Apoptosis Detection kit (Solarbio, China).

Techniques: Transfection, Western Blot, DCFH-DA Assay, Flow Cytometry

Journal: American Journal of Translational Research

Article Title: Knockdown of microRNA-96-5p resists oxidative stress-induced apoptosis in nucleus pulposus cells

doi:

Figure Lengend Snippet: MicroRNA-96-5p knockdown inhibited apoptosis in H2O2-treated NPCs. NPCs were transfected with inhibitor-NC or inhibitor-96-5p; cells were then treated with H2O2 (150 µM). A. Cell apoptosis was examined by flow cytometry. B. Protein levels of Bax, Bcl-2, and cleaved caspase3 were examined by western blot. **P<0.01.

Article Snippet: Apoptotic cells were examined by flow cytometric analysis (FCA) using an Annexin V-FITC Apoptosis Detection kit (Beyotime, China) according to the instructions.

Techniques: Knockdown, Transfection, Flow Cytometry, Western Blot

Journal: American Journal of Translational Research

Article Title: Knockdown of microRNA-96-5p resists oxidative stress-induced apoptosis in nucleus pulposus cells

doi:

Figure Lengend Snippet: MiR-298 knockdown regulated mitophagy depended on FOXO1 expression. A. NPCs were transfected with mimic-NC (negative control), mimic-96-5p, inhibitor-NC, or inhibitor-96-5p. Protein levels of FOXO1, PINK1, Parkin, and LC3 were examined by western blot. B. NPCs were transfected with mimic-NC (negative control), mimic-96-5p, or pCMV-FOXO1, cells were then treated with H2O2 (150 µM). Protein levels of FOXO1, PINK1, Parkin, and LC3 were examined by western blot. C. NPCs were transfected with mimic-NC (negative control), mimic-96-5p, or pCMV-FOXO1, and cells were then treated with H2O2 (150 µM). Protein levels of Bax, Bcl-2, and cleaved caspase3 were examined by western blot. D. NPCs were transfected with mimic-NC (negative control), mimic-96-5p, or pCMV-FOXO1, and cells were then treated with H2O2 (150 µM). Cell apoptosis was examined by flow cytometry. E. NPCs were transfected with mimic-NC (negative control), mimic-96-5p, or pCMV-FOXO1, cells were then treated with H2O2 (150 μM). Intracellular ROS production was measured by DCFH-DA assay. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Apoptotic cells were examined by flow cytometric analysis (FCA) using an Annexin V-FITC Apoptosis Detection kit (Beyotime, China) according to the instructions.

Techniques: Knockdown, Expressing, Transfection, Negative Control, Western Blot, Flow Cytometry, DCFH-DA Assay