Journal: Oncology reports

Article Title: NOB1 silencing inhibits the growth and metastasis of laryngeal cancer cells through the regulation of JNK signaling pathway.

doi: 10.3892/or.2016.4707

Figure Lengend Snippet: Figure 1. NOB1 level in laryngeal cancer patients. Paired laryngeal cancers and adjacent normal tissues were collected and the level of NOB1 was measured using (A) RT-qPCR and (B) western blotting. The results are presented as mean ± SD; *p<0.05, **p<0.01.

Article Snippet: After blockade with 5% skim milk or 1% BSA, the membranes were incubated with the primary antibodies against NOB1 (1:1,000; Proteintech, Chicago, IL, USA), cleaved-caspase-3, cleaved-PARP (1:1,000; Abcam, Cambridge, UK), matrix metalloproteinases (MMPs)-2, MMP-9, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax) (1:400; Boster, Wuhan, China), p-JNK, JNK (1:500; Bioss, Beijing, China) and β-actin (1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: Quantitative RT-PCR, Western Blot

Journal: Oncology reports

Article Title: NOB1 silencing inhibits the growth and metastasis of laryngeal cancer cells through the regulation of JNK signaling pathway.

doi: 10.3892/or.2016.4707

Figure Lengend Snippet: Figure 2. NOB1 shRNA downregulates the NOB1 level in laryngeal cancer cells. (A) After infection with NOB1 shRNA, the mRNA level of NOB1 was measured by RT-qPCR. (B and C) The protein level of NOB1 was detected by western blotting after infection with NOB1 shRNA. β-actin was used as the internal reference. Each experiment was repeated three times. Typical results are presented. The results are presented as mean ± SD; ***p<0.001.

Article Snippet: After blockade with 5% skim milk or 1% BSA, the membranes were incubated with the primary antibodies against NOB1 (1:1,000; Proteintech, Chicago, IL, USA), cleaved-caspase-3, cleaved-PARP (1:1,000; Abcam, Cambridge, UK), matrix metalloproteinases (MMPs)-2, MMP-9, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax) (1:400; Boster, Wuhan, China), p-JNK, JNK (1:500; Bioss, Beijing, China) and β-actin (1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: shRNA, Infection, Quantitative RT-PCR, Western Blot

Journal: Oncology reports

Article Title: NOB1 silencing inhibits the growth and metastasis of laryngeal cancer cells through the regulation of JNK signaling pathway.

doi: 10.3892/or.2016.4707

Figure Lengend Snippet: Figure 3. NOB1 shRNA inhibits the growth of laryngeal cancer cells. (A and B) Colony formation assay was performed after infection with NOB1 shRNA and the ratio of colony formation was calculated. (C) After infection with NOB1 shRNA, the cell viability was measured using MTT. (D and E) Cell cycle assay was performed after infection with NOB1 shRNA and the distribution of cell cycle was analyzed. All experiments were repeated three times and typical results are presented. Results are presented as mean ± SD; *p<0.05, **p<0.01.

Article Snippet: After blockade with 5% skim milk or 1% BSA, the membranes were incubated with the primary antibodies against NOB1 (1:1,000; Proteintech, Chicago, IL, USA), cleaved-caspase-3, cleaved-PARP (1:1,000; Abcam, Cambridge, UK), matrix metalloproteinases (MMPs)-2, MMP-9, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax) (1:400; Boster, Wuhan, China), p-JNK, JNK (1:500; Bioss, Beijing, China) and β-actin (1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: shRNA, Colony Assay, Infection, Cell Cycle Assay

Journal: Oncology reports

Article Title: NOB1 silencing inhibits the growth and metastasis of laryngeal cancer cells through the regulation of JNK signaling pathway.

doi: 10.3892/or.2016.4707

Figure Lengend Snippet: Figure 4. NOB1 silencing induces cell apoptosis. (A and B) Cell apoptosis was detected using flow cytometry and the percentage of cells in each phase was calculated. (C) Hoechst staining was used to detect cell apoptosis after infection with NOB1 shRNA. (D and E) Protein levels of cleaved PARP, cleaved caspase-3, Bax and Bcl-2 were detected using western blotting with β-actin as the internal reference. All experiments were repeated three times. Typical results are presented and results are presented as mean ± SD; **p<0.01, ***p<0.001.

Article Snippet: After blockade with 5% skim milk or 1% BSA, the membranes were incubated with the primary antibodies against NOB1 (1:1,000; Proteintech, Chicago, IL, USA), cleaved-caspase-3, cleaved-PARP (1:1,000; Abcam, Cambridge, UK), matrix metalloproteinases (MMPs)-2, MMP-9, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax) (1:400; Boster, Wuhan, China), p-JNK, JNK (1:500; Bioss, Beijing, China) and β-actin (1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: Flow Cytometry, Staining, Infection, shRNA, Western Blot

Journal: Oncology reports

Article Title: NOB1 silencing inhibits the growth and metastasis of laryngeal cancer cells through the regulation of JNK signaling pathway.

doi: 10.3892/or.2016.4707

Figure Lengend Snippet: Figure 5. Silencing of NOB1 inhibits migration of laryngeal cancer cells. (A) After infection with NOB1 shRNA, wound-healing assay was performed to detect changes in migration capacity. (B) Relative migration rate at 24 h. (C) Relative migration rate at 48 h. Each experiment was repeated three times and typical results are presented. Results are shown as mean ± SD; **p<0.01.

Article Snippet: After blockade with 5% skim milk or 1% BSA, the membranes were incubated with the primary antibodies against NOB1 (1:1,000; Proteintech, Chicago, IL, USA), cleaved-caspase-3, cleaved-PARP (1:1,000; Abcam, Cambridge, UK), matrix metalloproteinases (MMPs)-2, MMP-9, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax) (1:400; Boster, Wuhan, China), p-JNK, JNK (1:500; Bioss, Beijing, China) and β-actin (1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: Migration, Infection, shRNA, Wound Healing Assay

Journal: Oncology reports

Article Title: NOB1 silencing inhibits the growth and metastasis of laryngeal cancer cells through the regulation of JNK signaling pathway.

doi: 10.3892/or.2016.4707

Figure Lengend Snippet: Figure 6. Silencing of NOB1 inhibits invasion of laryngeal cancer cells. (A) Transwell assay was carried out after infection with NOB1 shRNA and the number of cells passing through the micropore membrane was calculated. (B and C) Protein levels of MMP-2 and MMP-9 were detected after infection with NOB1 shRNA. All experiments were repeated three times. Results are presented as mean ± SD and typical results are presented; **p<0.01, ***p<0.001.

Article Snippet: After blockade with 5% skim milk or 1% BSA, the membranes were incubated with the primary antibodies against NOB1 (1:1,000; Proteintech, Chicago, IL, USA), cleaved-caspase-3, cleaved-PARP (1:1,000; Abcam, Cambridge, UK), matrix metalloproteinases (MMPs)-2, MMP-9, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax) (1:400; Boster, Wuhan, China), p-JNK, JNK (1:500; Bioss, Beijing, China) and β-actin (1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: Transwell Assay, Infection, shRNA, Membrane

Journal: Oncology reports

Article Title: NOB1 silencing inhibits the growth and metastasis of laryngeal cancer cells through the regulation of JNK signaling pathway.

doi: 10.3892/or.2016.4707

Figure Lengend Snippet: Figure 7. NOB1 shRNA induces the activation of JNK. (A and B) Protein levels of JNK and phosphorylated JNK (p-JNK) were detected by western blotting. (C and D) Western blotting was used to detect the protein level of P38 and phosphorylated P38 (p-P38). Each experiment was repeated three times and the results are presented as mean ± SD. Typical results are presented; **p<0.01.

Article Snippet: After blockade with 5% skim milk or 1% BSA, the membranes were incubated with the primary antibodies against NOB1 (1:1,000; Proteintech, Chicago, IL, USA), cleaved-caspase-3, cleaved-PARP (1:1,000; Abcam, Cambridge, UK), matrix metalloproteinases (MMPs)-2, MMP-9, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax) (1:400; Boster, Wuhan, China), p-JNK, JNK (1:500; Bioss, Beijing, China) and β-actin (1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: shRNA, Activation Assay, Western Blot