primary antibodies against pdcd4  (Thermo Fisher)

Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-421-targeted PDCD4 regulates breast cancer cell proliferation

doi: 10.3892/ijmm.2018.3932

Figure Lengend Snippet: PDCD4 protein was detected in three sets of normal and breast cancer tissues using immunohistochemistry. (A) PDCD4 protein expression was detected in five normal and five BC tissues using immunohistochemistry, and decreased substantially in BC tissues compared with normal control tissues. (B) PDCD4 protein expression was determined by immunohistochemistry in five normal and five BC tissues, and was notably decreased in BC tissues. (C) PDCD4 protein expression was analyzed in five normal and five BC tissues by immunohistochemistry, but was downexpressed in BC tissues. BC, breast cancer; PDCD4, programmed cell death 4.

Article Snippet: The membranes were incubated with primary antibodies against PDCD4 (dilution 1:600; cat. no. PA5-20309; Invitrogen; Thermo Fisher Scientific, Inc.) and GAPDH (dilution 1:800; cat. no. MA5-15738; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 30 min.

Techniques: Immunohistochemistry, Expressing, Control

Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-421-targeted PDCD4 regulates breast cancer cell proliferation

doi: 10.3892/ijmm.2018.3932

Figure Lengend Snippet: Levels of miR-421 and expression of PDCD4 in BC. miR-421 levels were determined in BC tissues, MCF-7 cells and MDA-MB-231 cells, and the expression of PDCD4 was determined in MCF-7 cells and MDA-MB-231 cells. (A) Relative expression of miR-421 in 52 paired BC and normal tissues. (B) Relative expression of miR-421 in Hs578bst, MCF-7 and MDA-MB-231 cells. (C) Protein expression and PDCD4 in Hs578bst, MCF-7 and MDA-MB-231 cells was assessed using western blot analysis and (D) quantified. U6 was used as an endogenous control. * P<0.05 and ** P<0.01 vs. normal/control groups. BC, breast cancer; miR, microRNA; PDCD4, programmed cell death 4.

Article Snippet: The membranes were incubated with primary antibodies against PDCD4 (dilution 1:600; cat. no. PA5-20309; Invitrogen; Thermo Fisher Scientific, Inc.) and GAPDH (dilution 1:800; cat. no. MA5-15738; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 30 min.

Techniques: Expressing, Western Blot, Control

Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-421-targeted PDCD4 regulates breast cancer cell proliferation

doi: 10.3892/ijmm.2018.3932

Figure Lengend Snippet: PDCD4 is a direct target of miR-421. (A) A luciferase activity assay confirmed that miR-421 directly binds to PDCD4, as predicted by TargetScan. (B) Overexpression of miR-421 significantly inhibited the luciferase activity of PDCD4 in MCF-7 cells. (C) Luciferase activity of PDCD4 in MDA-MB-231 cells transfected with miR-421 inhibitors was significantly increased. * P<0.05 and ** P<0.01 vs. normal/control groups. miR, microRNA; Hum, human; PDCD4, programmed cell death 4; WT, wild-type; Mut, mutant.

Article Snippet: The membranes were incubated with primary antibodies against PDCD4 (dilution 1:600; cat. no. PA5-20309; Invitrogen; Thermo Fisher Scientific, Inc.) and GAPDH (dilution 1:800; cat. no. MA5-15738; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 30 min.

Techniques: Luciferase, Activity Assay, Over Expression, Transfection, Control, Mutagenesis

Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-421-targeted PDCD4 regulates breast cancer cell proliferation

doi: 10.3892/ijmm.2018.3932

Figure Lengend Snippet: Assessment of the expression of PDCD4 in breast cancer cell lines using RT-qPCR and western blot analysis. (A) miR-421 and (B) PDCD4 gene expression were measured in MCF-7 and MDA-MB-231 cells transfected with mock miRNA, miR-421 mimics and miR-421 inhibitors using RT-qPCR analysis. (C) Protein expression of PDCD4 was assessed in MCF-7 and MDA-MB-231 cells transfected with miR-421 mimics and miR-421 inhibitors using western blot analysis with (D) quantification of results. * P<0.05 and ** P<0.01 vs. normal/control groups. miR, microRNA; PDCD4, programmed cell death 4; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.

Article Snippet: The membranes were incubated with primary antibodies against PDCD4 (dilution 1:600; cat. no. PA5-20309; Invitrogen; Thermo Fisher Scientific, Inc.) and GAPDH (dilution 1:800; cat. no. MA5-15738; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 30 min.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Gene Expression, Transfection, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Therapy. Nucleic Acids

Article Title: Identifying Involvement of H19-miR-675-3p-IGF1R and H19-miR-200a-PDCD4 in Treating Pulmonary Hypertension with Melatonin

doi: 10.1016/j.omtn.2018.08.015

Figure Lengend Snippet: Differential Levels of H19, miR-675, miR-200a, IGF1R, and PDCD4 after MCT Treatment in PAH Rats (A) The H19 level in the MCT plus melatonin group was much higher than in the MCT group, while it was even higher in the control group than in the MCT plus melatonin group (*p < 0.05 as compared with the control group, #p < 0.05 as compared with the MCT group). (B) The miR-200a level in the MCT plus melatonin group was much higher than in the control group, while it was even higher in the MCT group than in the MCT plus melatonin group (*p < 0.05 as compared with the control group, #p < 0.05 as compared with the MCT group). (C) The miR-675-3p level in the MCT group was much lower than in the MCT plus melatonin group, and both of them were much lower than in the control group (*p < 0.05 as compared with the control group, #p < 0.05 as compared with the MCT group). (D) The IGF1R mRNA level in the MCT group was much higher than in the MCT plus melatonin group, and both of them were much higher than in the control group (*p < 0.05 as compared with the control group, #p < 0.05 as compared with the MCT group). (E) The MCT plus melatonin group displayed a lower level of PDCD4 mRNA than did the control group, while the MCT group exhibited an even lower level of PDCD4 mRNA than did the MCT plus melatonin group (*p < 0.05 as compared with the control group, #p < 0.05 as compared with the MCT group). (F) The IGF1R protein level in the MCT group was much higher than in the MCT plus melatonin group, and both of them were much higher than in the control group. The PDCD4 protein level in the MCT group was much lower than in the MCT plus melatonin group, and both of them were much lower than in the control group (*p < 0.05 as compared with the control group, #p < 0.05 as compared with the MCT group).

Article Snippet: Subsequently, the sections were blocked with 3% H 2 O 2 to inactivate the activity of endogenous peroxidase, followed by incubation at room temperature for 2 hr with primary goat antibodies against PDCD4 (1:200 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and IGF1R (1:200 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: Identifying Involvement of H19-miR-675-3p-IGF1R and H19-miR-200a-PDCD4 in Treating Pulmonary Hypertension with Melatonin

doi: 10.1016/j.omtn.2018.08.015

Figure Lengend Snippet: Immunohistochemistry for PDCD4 According to the IHC result, the PDCD4 protein level in the MCT group was much lower than in the MCT plus melatonin group, and both of them were much lower than in the control group (scale bars, 10 μm).

Article Snippet: Subsequently, the sections were blocked with 3% H 2 O 2 to inactivate the activity of endogenous peroxidase, followed by incubation at room temperature for 2 hr with primary goat antibodies against PDCD4 (1:200 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and IGF1R (1:200 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Immunohistochemistry, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: Identifying Involvement of H19-miR-675-3p-IGF1R and H19-miR-200a-PDCD4 in Treating Pulmonary Hypertension with Melatonin

doi: 10.1016/j.omtn.2018.08.015

Figure Lengend Snippet: miR-675-3p and miR-200a Directly Targeted IGF1R and PDCD4, Respectively (A) The sequence comparison between mature miR-675-3p and wild-type as well as mutant IGF1R 3′ UTR. (B) Luciferase activity of wild-type IGF1R 3′ UTR, but not that of mutant IGF1R 3′ UTR, in hPASMCs transfected with miR-675-3p mimic was much lower than scramble control (*p < 0.05 as compared with the control group). (C) Luciferase activity of wild-type IGF1R 3′ UTR, but not that of mutant IGF1R 3′ UTR, in rPASMCs transfected with miR-675 mimic was much lower than scramble control (*p < 0.05 as compared with the control group). (D) The sequence comparison between mature miR-200a and wild-type as well as mutant PDCD4 3′ UTR. (E) Luciferase activity of wild-type PDCD4 3′ UTR, but not that of mutant PDCD4 3′ UTR, in hPASMCs transfected with miR-675 mimic was downregulated compared with scramble control (*p < 0.05 as compared with the control group). (F) Transfecting with miR-200a mimic reduced luciferase activity of wild-type PDCD4 3′ UTR, but not that of mutant PDCD4 3′ UTR, in rPASMCs (*p < 0.05 as compared with the control group).

Article Snippet: Subsequently, the sections were blocked with 3% H 2 O 2 to inactivate the activity of endogenous peroxidase, followed by incubation at room temperature for 2 hr with primary goat antibodies against PDCD4 (1:200 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and IGF1R (1:200 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Sequencing, Comparison, Mutagenesis, Luciferase, Activity Assay, Transfection, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: Identifying Involvement of H19-miR-675-3p-IGF1R and H19-miR-200a-PDCD4 in Treating Pulmonary Hypertension with Melatonin

doi: 10.1016/j.omtn.2018.08.015

Figure Lengend Snippet: Effect of Melatonin on Cell Proliferation and H19, miR-675-3p, miR-200a, IGF1R, and PDCD4 Levels in hPASMCs (A) Melatonin inhibited cell viability of hPASMCs in a dose-dependent manner (*p < 0.05 as compared with the control group). (B) Melatonin inhibited cell viability of hPASMCs in a dose-dependent fashion. (C) Treating with melatonin dose-dependently upregulated H19 expression (*p < 0.05 as compared with the control group). (D) Treating with melatonin dose-dependently enhanced miR-675-3p expression (*p < 0.05 as compared with the control group). (E) miR-200a level was dose-dependently reduced following treatment with melatonin (*p < 0.05 as compared with the control group). (F) IGF1R mRNA level was dose-dependently suppressed after the administration of melatonin (*p < 0.05 as compared with the control group). (G) PDCD4 mRNA level was dose-dependently upregulated subsequent to treatment with melatonin (*p < 0.05 as compared with the control group). (H) IGF1R protein level was dose-dependently suppressed while PDCD4 protein expression was dose-dependently enhanced following treatment with melatonin.

Article Snippet: Subsequently, the sections were blocked with 3% H 2 O 2 to inactivate the activity of endogenous peroxidase, followed by incubation at room temperature for 2 hr with primary goat antibodies against PDCD4 (1:200 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and IGF1R (1:200 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Control, Expressing

Journal: Molecular Therapy. Nucleic Acids

Article Title: Identifying Involvement of H19-miR-675-3p-IGF1R and H19-miR-200a-PDCD4 in Treating Pulmonary Hypertension with Melatonin

doi: 10.1016/j.omtn.2018.08.015

Figure Lengend Snippet: Effect of Melatonin on Cell Proliferation and H19, miR-675, miR-200a, IGF1R, and PDCD4 Levels in rPASMCs (A) Melatonin inhibited cell viability of rPASMCs in a dose-dependent manner (*p < 0.05 as compared with the control group). (B) Melatonin inhibited cell viability of rPASMCs in a dose-dependent fashion. (C) Treating with melatonin dose-dependently upregulated H19 expression (*p < 0.05 as compared with the control group). (D) Treating with melatonin dose-dependently enhanced miR-675-3p expression (*p < 0.05 as compared with the control group). (E) miR-200a level was dose-dependently reduced following treatment with melatonin (*p < 0.05 as compared with the control group). (F) IGF1R mRNA level was dose-dependently suppressed after the administration of melatonin (*p < 0.05 as compared with the control group). (G) PDCD4 mRNA level was dose-dependently upregulated subsequent to treatment with melatonin (*p < 0.05 as compared with the control group). (H) IGF1R protein level was dose-dependently suppressed while PDCD4 protein expression was dose-dependently enhanced following treatment with melatonin.

Article Snippet: Subsequently, the sections were blocked with 3% H 2 O 2 to inactivate the activity of endogenous peroxidase, followed by incubation at room temperature for 2 hr with primary goat antibodies against PDCD4 (1:200 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and IGF1R (1:200 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Control, Expressing

Journal: Molecular Therapy. Nucleic Acids

Article Title: Identifying Involvement of H19-miR-675-3p-IGF1R and H19-miR-200a-PDCD4 in Treating Pulmonary Hypertension with Melatonin

doi: 10.1016/j.omtn.2018.08.015

Figure Lengend Snippet: Two Signaling Pathways Are Shown Dual signaling pathways (H19-miR-675-3p-IGF1R and H19-miR-200a-PDCD4) were involved in the mechanisms underlying the therapeutic effect of melatonin in the treatment of PAH by promoting the apoptosis of PASMCs.

Article Snippet: Subsequently, the sections were blocked with 3% H 2 O 2 to inactivate the activity of endogenous peroxidase, followed by incubation at room temperature for 2 hr with primary goat antibodies against PDCD4 (1:200 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and IGF1R (1:200 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Protein-Protein interactions

Journal: Molecular Therapy. Nucleic Acids

Article Title: Identifying Involvement of H19-miR-675-3p-IGF1R and H19-miR-200a-PDCD4 in Treating Pulmonary Hypertension with Melatonin

doi: 10.1016/j.omtn.2018.08.015

Figure Lengend Snippet: Differential Levels of H19, miR-675, miR-200a, IGF1R, and PDCD4 after MCT Treatment in PAH Rats (A) The H19 level in the MCT plus melatonin group was much higher than in the MCT group, while it was even higher in the control group than in the MCT plus melatonin group (*p < 0.05 as compared with the control group, #p < 0.05 as compared with the MCT group). (B) The miR-200a level in the MCT plus melatonin group was much higher than in the control group, while it was even higher in the MCT group than in the MCT plus melatonin group (*p < 0.05 as compared with the control group, #p < 0.05 as compared with the MCT group). (C) The miR-675-3p level in the MCT group was much lower than in the MCT plus melatonin group, and both of them were much lower than in the control group (*p < 0.05 as compared with the control group, #p < 0.05 as compared with the MCT group). (D) The IGF1R mRNA level in the MCT group was much higher than in the MCT plus melatonin group, and both of them were much higher than in the control group (*p < 0.05 as compared with the control group, #p < 0.05 as compared with the MCT group). (E) The MCT plus melatonin group displayed a lower level of PDCD4 mRNA than did the control group, while the MCT group exhibited an even lower level of PDCD4 mRNA than did the MCT plus melatonin group (*p < 0.05 as compared with the control group, #p < 0.05 as compared with the MCT group). (F) The IGF1R protein level in the MCT group was much higher than in the MCT plus melatonin group, and both of them were much higher than in the control group. The PDCD4 protein level in the MCT group was much lower than in the MCT plus melatonin group, and both of them were much lower than in the control group (*p < 0.05 as compared with the control group, #p < 0.05 as compared with the MCT group).

Article Snippet: Subsequently, the membrane was incubated at 4°C for 12 hr with anti-β-actin monoclonal antibodies (1:8,000 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA) or polyclonal goat primary antibodies against PDCD4 and IGF1R (1:5,000 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: Identifying Involvement of H19-miR-675-3p-IGF1R and H19-miR-200a-PDCD4 in Treating Pulmonary Hypertension with Melatonin

doi: 10.1016/j.omtn.2018.08.015

Figure Lengend Snippet: Immunohistochemistry for IGF1R According to the IHC result, the IGF1R protein level in the MCT group was much higher than in the MCT plus melatonin group, and both of them were much higher than in the control group (scale bars, 10 μm).

Article Snippet: Subsequently, the membrane was incubated at 4°C for 12 hr with anti-β-actin monoclonal antibodies (1:8,000 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA) or polyclonal goat primary antibodies against PDCD4 and IGF1R (1:5,000 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Immunohistochemistry, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: Identifying Involvement of H19-miR-675-3p-IGF1R and H19-miR-200a-PDCD4 in Treating Pulmonary Hypertension with Melatonin

doi: 10.1016/j.omtn.2018.08.015

Figure Lengend Snippet: miR-675-3p and miR-200a Directly Targeted IGF1R and PDCD4, Respectively (A) The sequence comparison between mature miR-675-3p and wild-type as well as mutant IGF1R 3′ UTR. (B) Luciferase activity of wild-type IGF1R 3′ UTR, but not that of mutant IGF1R 3′ UTR, in hPASMCs transfected with miR-675-3p mimic was much lower than scramble control (*p < 0.05 as compared with the control group). (C) Luciferase activity of wild-type IGF1R 3′ UTR, but not that of mutant IGF1R 3′ UTR, in rPASMCs transfected with miR-675 mimic was much lower than scramble control (*p < 0.05 as compared with the control group). (D) The sequence comparison between mature miR-200a and wild-type as well as mutant PDCD4 3′ UTR. (E) Luciferase activity of wild-type PDCD4 3′ UTR, but not that of mutant PDCD4 3′ UTR, in hPASMCs transfected with miR-675 mimic was downregulated compared with scramble control (*p < 0.05 as compared with the control group). (F) Transfecting with miR-200a mimic reduced luciferase activity of wild-type PDCD4 3′ UTR, but not that of mutant PDCD4 3′ UTR, in rPASMCs (*p < 0.05 as compared with the control group).

Article Snippet: Subsequently, the membrane was incubated at 4°C for 12 hr with anti-β-actin monoclonal antibodies (1:8,000 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA) or polyclonal goat primary antibodies against PDCD4 and IGF1R (1:5,000 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Sequencing, Comparison, Mutagenesis, Luciferase, Activity Assay, Transfection, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: Identifying Involvement of H19-miR-675-3p-IGF1R and H19-miR-200a-PDCD4 in Treating Pulmonary Hypertension with Melatonin

doi: 10.1016/j.omtn.2018.08.015

Figure Lengend Snippet: Effect of Melatonin on Cell Proliferation and H19, miR-675-3p, miR-200a, IGF1R, and PDCD4 Levels in hPASMCs (A) Melatonin inhibited cell viability of hPASMCs in a dose-dependent manner (*p < 0.05 as compared with the control group). (B) Melatonin inhibited cell viability of hPASMCs in a dose-dependent fashion. (C) Treating with melatonin dose-dependently upregulated H19 expression (*p < 0.05 as compared with the control group). (D) Treating with melatonin dose-dependently enhanced miR-675-3p expression (*p < 0.05 as compared with the control group). (E) miR-200a level was dose-dependently reduced following treatment with melatonin (*p < 0.05 as compared with the control group). (F) IGF1R mRNA level was dose-dependently suppressed after the administration of melatonin (*p < 0.05 as compared with the control group). (G) PDCD4 mRNA level was dose-dependently upregulated subsequent to treatment with melatonin (*p < 0.05 as compared with the control group). (H) IGF1R protein level was dose-dependently suppressed while PDCD4 protein expression was dose-dependently enhanced following treatment with melatonin.

Article Snippet: Subsequently, the membrane was incubated at 4°C for 12 hr with anti-β-actin monoclonal antibodies (1:8,000 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA) or polyclonal goat primary antibodies against PDCD4 and IGF1R (1:5,000 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Control, Expressing

Journal: Molecular Therapy. Nucleic Acids

Article Title: Identifying Involvement of H19-miR-675-3p-IGF1R and H19-miR-200a-PDCD4 in Treating Pulmonary Hypertension with Melatonin

doi: 10.1016/j.omtn.2018.08.015

Figure Lengend Snippet: Effect of Melatonin on Cell Proliferation and H19, miR-675, miR-200a, IGF1R, and PDCD4 Levels in rPASMCs (A) Melatonin inhibited cell viability of rPASMCs in a dose-dependent manner (*p < 0.05 as compared with the control group). (B) Melatonin inhibited cell viability of rPASMCs in a dose-dependent fashion. (C) Treating with melatonin dose-dependently upregulated H19 expression (*p < 0.05 as compared with the control group). (D) Treating with melatonin dose-dependently enhanced miR-675-3p expression (*p < 0.05 as compared with the control group). (E) miR-200a level was dose-dependently reduced following treatment with melatonin (*p < 0.05 as compared with the control group). (F) IGF1R mRNA level was dose-dependently suppressed after the administration of melatonin (*p < 0.05 as compared with the control group). (G) PDCD4 mRNA level was dose-dependently upregulated subsequent to treatment with melatonin (*p < 0.05 as compared with the control group). (H) IGF1R protein level was dose-dependently suppressed while PDCD4 protein expression was dose-dependently enhanced following treatment with melatonin.

Article Snippet: Subsequently, the membrane was incubated at 4°C for 12 hr with anti-β-actin monoclonal antibodies (1:8,000 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA) or polyclonal goat primary antibodies against PDCD4 and IGF1R (1:5,000 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Control, Expressing

Journal: Molecular Therapy. Nucleic Acids

Article Title: Identifying Involvement of H19-miR-675-3p-IGF1R and H19-miR-200a-PDCD4 in Treating Pulmonary Hypertension with Melatonin

doi: 10.1016/j.omtn.2018.08.015

Figure Lengend Snippet: Two Signaling Pathways Are Shown Dual signaling pathways (H19-miR-675-3p-IGF1R and H19-miR-200a-PDCD4) were involved in the mechanisms underlying the therapeutic effect of melatonin in the treatment of PAH by promoting the apoptosis of PASMCs.

Article Snippet: Subsequently, the membrane was incubated at 4°C for 12 hr with anti-β-actin monoclonal antibodies (1:8,000 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA) or polyclonal goat primary antibodies against PDCD4 and IGF1R (1:5,000 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Protein-Protein interactions

Journal: Molecular Medicine Reports

Article Title: MicroRNA-206 contributes to the progression of steroid-induced avascular necrosis of the femoral head by inducing osteoblast apoptosis by suppressing programmed cell death 4

doi: 10.3892/mmr.2017.7963

Figure Lengend Snippet: Analysis of the expression of miR-206 and PDCD4 in the SANFH specimens. Investigation using RT-qPCR demonstrated that the (A) mRNA expression level of miR-206 was increased compared with the control, while (B) RT-qPCR and (C) western blotting demonstrated that PDCD4 was inhibited in SANFH specimens compared with the control. (D) Correlation analysis between the relative expression of PDCDA and miR-206. There was a negative correlation between the expression levels of the two indicators. Data are presented as the mean ± standard deviation. **P<0.01 and ***P<0.001 vs. control. RT-qPCR, reverse transcription-quantitative polymerase chain reaction; miR, microRNA; PDCD4, programmed cell death 4; SANFH, steroid-induced avascular necrosis of the femoral head.

Article Snippet: Following blocking in 10% goat serum (Thermo Fisher Scientific, Inc.) for 15 min, primary rabbit polyclonal antibodies against different proteins of interest (PDCD4) (1:500) were then added, and the cells were incubated overnight at 4°C.

Techniques: Expressing, Quantitative RT-PCR, Control, Western Blot, Standard Deviation, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: MicroRNA-206 contributes to the progression of steroid-induced avascular necrosis of the femoral head by inducing osteoblast apoptosis by suppressing programmed cell death 4

doi: 10.3892/mmr.2017.7963

Figure Lengend Snippet: Complementary sequences of hsa-miR-206 and PDCD4 and results of a dual luciferase assay demonstrating direct binding of miR-206 to the promoter sequence of PDCD4 gene. Data are presented as the mean ± standard deviation. **P<0.01 vs. NC. NC, negative control; miR, microRNA; PDCD4, programmed cell death 4; WT, wild type; Mut, mutant; UTR, untranslated region. R/F, Renilla to firefly activity ratio.

Article Snippet: Following blocking in 10% goat serum (Thermo Fisher Scientific, Inc.) for 15 min, primary rabbit polyclonal antibodies against different proteins of interest (PDCD4) (1:500) were then added, and the cells were incubated overnight at 4°C.

Techniques: Luciferase, Binding Assay, Sequencing, Standard Deviation, Negative Control, Mutagenesis, Activity Assay

Journal: Molecular Medicine Reports

Article Title: MicroRNA-206 contributes to the progression of steroid-induced avascular necrosis of the femoral head by inducing osteoblast apoptosis by suppressing programmed cell death 4

doi: 10.3892/mmr.2017.7963

Figure Lengend Snippet: Regulation of miR-206 influences the expression of the different signaling molecules associated with differentiation and apoptosis in hFOB1.19 cells. (A) Relative expression of miR-206, PDCD4 and ALP in hFOB1.19 cells transfected with an miR-206 inhibitor, mimic or NC. (B) Western blotting demonstrating the expression of PDCD4, ALP, Bcl-2, Bax and GAPDH in hFOB1.19 cells transfected with an miR-206 inhibitor, mimic or NC. (C) Transfection of miR-206 mimic suppressed the expression of PDCD4. (D) Transfection of miR-206 inhibitor induced the expression of PDCD4. *P<0.05, **P<0.01 vs. NC. ##P<0.01 vs. mimic. PDCD4, programmed cell death 4; ALP, alkaline phosphatase; Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-X-associated protein; miR, microRNA; NC, negative control; DAPI, 4,6-diamino-2-phenylindole.

Article Snippet: Following blocking in 10% goat serum (Thermo Fisher Scientific, Inc.) for 15 min, primary rabbit polyclonal antibodies against different proteins of interest (PDCD4) (1:500) were then added, and the cells were incubated overnight at 4°C.

Techniques: Expressing, Transfection, Western Blot, Negative Control

Journal: Molecular Medicine Reports

Article Title: MicroRNA-206 contributes to the progression of steroid-induced avascular necrosis of the femoral head by inducing osteoblast apoptosis by suppressing programmed cell death 4

doi: 10.3892/mmr.2017.7963

Figure Lengend Snippet: Analysis of the effect of miR-206 on the differentiation potential and apoptosis in hFOB1.19 cells depended on the suppression of PDCD4 associated signaling. Analysis of the relative expression of (A) PDCD4 and ALP in hFOB1.19 cells transfected with the miR-206 inhibitor and an siRNA against PDCD4. (B) Cell apoptosis and quantification, detected by flow cytometry of cells transfected with miR-206 inhibitor and an siRNA against PDCD4. (C) Western blotting and quantification demonstrating the expression of PDCD4, ALP, Bcl-2 and Bax, in hFOB1.19 cells transfected with miR-206 inhibitor and an siRNA against PDCD4. (D) Hoechest staining of hFOB1.19 cells transfected with the miR-206 inhibitor and an siRNA against PDCD4. Data are expressed as the mean ± standard deviation. *P<0.05, **P<0.01, ***P<0.001 vs. miR-206 inhibited hFOB1.19 cells. NC, negative control; PDCD4, programmed cell death 4; ALP, alkaline phosphatase; Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-X-associated protein; miR, microRNA; si, small interfering; FITC, fluorescein isothiocyanate; PI, propidium iodide.

Article Snippet: Following blocking in 10% goat serum (Thermo Fisher Scientific, Inc.) for 15 min, primary rabbit polyclonal antibodies against different proteins of interest (PDCD4) (1:500) were then added, and the cells were incubated overnight at 4°C.

Techniques: Expressing, Transfection, Flow Cytometry, Western Blot, Staining, Standard Deviation, Negative Control