Journal: Oncotarget

Article Title: Multiple-level validation identifies PARK2 in the development of lung cancer and chronic obstructive pulmonary disease

doi: 10.18632/oncotarget.9954

Figure Lengend Snippet: A-C. Cytokines, IL-6, IL-1β or TNF-α protein or mRNA levels in serum from C57BL/6 mice (A), mouse primary bronchus epithelial cells (B), MEFs (C) of PARK2 WT or KO were analyzed by ELISA or qRT-PCR. D. After infection with the indicated plasmids, cytokines in the cultured MEFs supernatants were determined by ELISA. Results are shown as means (±SEM), and at least three experiments were performed for all experiments. ***, p < 0.001 by one-way ANOVA. E. PARK2 WT and KO mice ( n=6 ) at 10 months of age were analyzed for tumor event by immunohistochemistry. H&E staining; x100 and x400.

Article Snippet: The pMX retroviral vector containing the human cDNAs for HA-PARK2 was obtained from Addgene (plasmid 38248, Cambridge, MA).

Techniques: Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Infection, Cell Culture, Immunohistochemistry, Staining

Journal: Oncotarget

Article Title: Multiple-level validation identifies PARK2 in the development of lung cancer and chronic obstructive pulmonary disease

doi: 10.18632/oncotarget.9954

Figure Lengend Snippet: A. PARK2 deficiency results in centrosome amplification in MEFs at passage 5. ( left ) Immunofluorescence staining shows impaired mitoses in MEFs depleted of PARK2 . Red, γ-tubulin; White arrows, centrosome in mitosis. The scale bar is 20 μm. ( right ) Quantification of 2 < centrosomes cells. ***, p < 0.001 by one-way ANOVA. n =10. B. After infection with the indicated plasmids, MEFs lysates were fixed for immunofluorescence assay. Quantification of 2 < centrosomes cells using γ-tubulin. Results are shown as means (±SEM), and at least three experiments were performed for all experiments. *, p < 0.05 and **, p < 0.01 by one-way ANOVA. C. Immunofluorescence staining shows impaired mitoses in IMR 90 lung fibroblast cells depleted of PARK2 . White arrows, centrosome in mitosis. Yellow arrows, abnormal chromosome. The scale bar is 20 μm. D. Cell were harvested at indicated times and analyzed by immunobloting. E. Impaired mitoses including multipolar spindles, misalignment and abnormal microtubule in PARK2 knockout (KO) MEF cells. Yellow arrows, abnormal chromosome. The scale bar is 20 μm. Results are shown as means (±SEM), and at least three experiments were performed for all experiments. **, p < 0.01 by one-way ANOVA.

Article Snippet: The pMX retroviral vector containing the human cDNAs for HA-PARK2 was obtained from Addgene (plasmid 38248, Cambridge, MA).

Techniques: Amplification, Immunofluorescence, Staining, Infection, Western Blot, Knock-Out

Journal: Oncotarget

Article Title: Multiple-level validation identifies PARK2 in the development of lung cancer and chronic obstructive pulmonary disease

doi: 10.18632/oncotarget.9954

Figure Lengend Snippet: A. Primary MEFs were applied on 3-D cultures system for 2 weeks and counted the sphere of the cells. ( top ) Representative images indicate the sphere cells. ( bottom ) Quantification of the sphere number. ****, p < 0.0001 by one-way ANOVA. n =6. ( B. and C. ) PARK2 WT and KO MEFs were analyzed for foci formation at passage 5 and 21 (B), and colony formation at passage 21 C. The scale bar is 20 μm. Results are shown as means (±SEM), and at least three experiments were performed for all experiments. NAC, ROS scavenger. ***, p < 0.001 by one-way ANOVA. D. The growth of lung cancer cell lines (H1437, H522, H1650, A549, H460, H1299 and H196) was inhibited through infection with PARK2 . Cells were transfected with the indicated plasmids, and then cell lysates were blotted with the indicated antibodies. E. The percentage of lung cancer cell growth compared to controls after transfected PARK2 . Results shown as means (± SEM), and at least three experiments were performed for all experiments. **, p < 0.01 and ***, p < 0.001 by one-way ANOVA. F. PARK2 and β–actin expression was measured by immunoblot.

Article Snippet: The pMX retroviral vector containing the human cDNAs for HA-PARK2 was obtained from Addgene (plasmid 38248, Cambridge, MA).

Techniques: Infection, Transfection, Expressing, Western Blot

Journal: Oncotarget

Article Title: Multiple-level validation identifies PARK2 in the development of lung cancer and chronic obstructive pulmonary disease

doi: 10.18632/oncotarget.9954

Figure Lengend Snippet: Significant results of Testing 114 Informative PARK2 SNPs

Article Snippet: The pMX retroviral vector containing the human cDNAs for HA-PARK2 was obtained from Addgene (plasmid 38248, Cambridge, MA).

Techniques: