park2  (Addgene inc)

Journal: International Journal of Molecular Sciences

Article Title: Parkin Precipitates on Mitochondria via Aggregation and Autoubiquitination

doi: 10.3390/ijms24109027

Figure Lengend Snippet: List of plasmids.

Article Snippet: pCMV3-non-tagged Parkin , Sino Biological, Beijing, PR China , HG12092-UT.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Parkin Precipitates on Mitochondria via Aggregation and Autoubiquitination

doi: 10.3390/ijms24109027

Figure Lengend Snippet: List of plasmids.

Article Snippet: pCMV3-non-tagged Parkin , Sino Biological, Beijing, PR China , HG12092-UT.

Techniques:

Journal: Molecules and Cells

Article Title: PARK2 Induces Osteoclastogenesis through Activation of the NF-κB Pathway

doi: 10.14348/molcells.2022.0058

Figure Lengend Snippet: (A) The mRNA levels of PARK2 and osteoclast marker genes were analyzed by real-time PCR in BMMs cultured with RANKL and M-CSF for the indicated days. *** P < 0.001 versus day 0. (B) PARK2 protein expression during osteoclast differentiation were analyzed by western blotting. (C) The relative level of PARK2 protein was determined by Image J software. *** P < 0.001 versus BMM. Data were presented as mean ± SD.

Article Snippet: PARK2 plasmid (#59416) was purchased from Addgene (USA).

Techniques: Marker, Real-time Polymerase Chain Reaction, Cell Culture, Expressing, Western Blot, Software

Journal: Molecules and Cells

Article Title: PARK2 Induces Osteoclastogenesis through Activation of the NF-κB Pathway

doi: 10.14348/molcells.2022.0058

Figure Lengend Snippet: (A) pOCs transfected with control (Con) or PARK2 siRNA were subjected to western blotting and real-time PCR analyses. * P < 0.05 versus the control siRNA group. (B) pOCs transfected with control or PARK2 siRNA were further cultured with RANKL and M-CSF for 3 days and then stained for TRAP. TRAP + multinucleated cells more than 3, 10 nuclei were counted. Scale bars = 200 μm. * P < 0.05, *** P < 0.001. (C) pOCs transfected with control or PARK2 siRNA were cultured with RANKL and M-CSF until day 6. The mRNA expression levels of ACP5, DC-STMAP, MMP9 and v-ATPase were analyzed by real-time PCR. * P < 0.05; ** P < 0.01; *** P < 0.001. Data were presented as mean ± SD. (D) pOCs transfected with control or PARK2 siRNA were cultured on dentine discs until day 12. The dark areas in gray images indicate the resorbed surface of dentin slices. The percentage of resorbed area and depth of resorption pits were presented as mean ± SD. * P < 0.05; ** P < 0.01.

Article Snippet: PARK2 plasmid (#59416) was purchased from Addgene (USA).

Techniques: Transfection, Control, Western Blot, Real-time Polymerase Chain Reaction, Cell Culture, Staining, Expressing

Journal: Molecules and Cells

Article Title: PARK2 Induces Osteoclastogenesis through Activation of the NF-κB Pathway

doi: 10.14348/molcells.2022.0058

Figure Lengend Snippet: (A) BMMs transfected with control (Con) vector or PARK2 plasmid were subjected to western blotting (left panel) and real-time PCR (right panel) analyses. *** P < 0.001. (B) BMMs transfected with either control or PARK2 plasmid were cultured with RANKL and M-CSF for 5 days and stained for TRAP. Representative images (left panel) and quantification of TRAP + multinucleated cells are shown (right panel). Scale bars = 200 μm. *** P < 0.001. (C) mRNA levels of ACP5, DC-STAMP, MMP9 and OSCAR were measured by real-time PCR. * P < 0.05; *** P < 0.001.

Article Snippet: PARK2 plasmid (#59416) was purchased from Addgene (USA).

Techniques: Transfection, Control, Plasmid Preparation, Western Blot, Real-time Polymerase Chain Reaction, Cell Culture, Staining

Journal: Molecules and Cells

Article Title: PARK2 Induces Osteoclastogenesis through Activation of the NF-κB Pathway

doi: 10.14348/molcells.2022.0058

Figure Lengend Snippet: (A and B) pOCs transfected with control (Con) siRNA or PARK2 siRNA were starved with serum-free medium and then stimulated with RANKL (500 ng/ml) for the indicated time. Both phospho-forms and total forms of the NF-κB signaling components (A) and MAPKs (B) were determined by western blotting. (C) pOCs transfected with control siRNA or PARK2 siRNA were serum-starved and then stimulated with RANKL (500 ng/ml) for 15 min. Representative confocal images are presented. Scale bars = 20 μm. *** P < 0.001. (D) BMMs transfected with control or PARK2 plasmid were cultured with or without BAY 11-7085 (5 μM) for 2 days. Protein levels of phospho-forms and total forms of p65 were detected by western blotting.

Article Snippet: PARK2 plasmid (#59416) was purchased from Addgene (USA).

Techniques: Transfection, Control, Western Blot, Plasmid Preparation, Cell Culture

Journal: Molecules and Cells

Article Title: PARK2 Induces Osteoclastogenesis through Activation of the NF-κB Pathway

doi: 10.14348/molcells.2022.0058

Figure Lengend Snippet: (A) BMMs were cultured in the presence of LPS (100 ng/ml), IL-1β (10 ng/ml), and TNFα (40 ng/ml) for 24 h and mRNA levels of PARK2 were determined by real-time PCR. ** P < 0.01; *** P < 0.001. Con, control. (B) pOCs transfected with control siRNA or PARK2 siRNA were cultured in the presence of LPS (100 ng/ml), IL-1β (10 ng/ml), or TNFα (40 ng/ml) with RANKL and M-CSF until day 4 and stained for TRAP. Scale bars = 200 μm. (C) mRNA levels of ACP5, DC-STAMP, OC-STAMP, and v-ATPase in PARK2 or control siRNA transfected pOC cultured with RANKL and LPS (100 ng/ml) were measured by real-time PCR. * P < 0.05; ** P < 0.01; *** P < 0.001. (D) pOCs transfected with control siRNA or PARK2 siRNA were serum-starved and stimulated with LPS (200 ng/ml) for 15 min. Protein levels of the phospho-forms and total forms of IKK, IκB, and p65 were determined by western blotting.

Article Snippet: PARK2 plasmid (#59416) was purchased from Addgene (USA).

Techniques: Cell Culture, Real-time Polymerase Chain Reaction, Control, Transfection, Staining, Western Blot

Journal: Molecules and Cells

Article Title: PARK2 Induces Osteoclastogenesis through Activation of the NF-κB Pathway

doi: 10.14348/molcells.2022.0058

Figure Lengend Snippet: (A) LPS (2.5 mg/kg) or vehicle (PBS) was injected at the midline of calvariae. Either control (Con) or PARK2 siRNA was locally injected near the LPS-injected site. The knock-down efficiency was determined by real-time PCR. * P < 0.05; *** P < 0.001. (B) Representative μCT images (left panel) and measurements of bone volume per tissue volume (BV/TV) (right panel) are shown. * P < 0.05; *** P < 0.001. (C) Total calvariae were stained for TRAP activity (left panel). Quantitative values of TRAP + area (%) are shown at the right panel. * P < 0.05; *** P < 0.001.

Article Snippet: PARK2 plasmid (#59416) was purchased from Addgene (USA).

Techniques: Injection, Control, Knockdown, Real-time Polymerase Chain Reaction, Staining, Activity Assay

Journal: Molecules and Cells

Article Title: PARK2 Induces Osteoclastogenesis through Activation of the NF-κB Pathway

doi: 10.14348/molcells.2022.0058

Figure Lengend Snippet: Upon activation of RANK by RANKL, TRAF6 is recruited and activated. Downstream of TRAF6, PARK2 may interact with NEMO and enhance its linear ubiquitination, resulting in the activation of the IKK complex. The activated IKK complex facilitates proteasomal degradation of IκB and triggers p65 translocation to the nucleus, leading to gene transcription of osteoclast marker genes like ACP5, MMP9, and Ctsk.

Article Snippet: PARK2 plasmid (#59416) was purchased from Addgene (USA).

Techniques: Activation Assay, Ubiquitin Proteomics, Translocation Assay, Marker