parkin  (Proteintech)

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: Targeting VCP with V8 suppresses glioblastoma development via formation of aggregates and disruption of mitophagy flux

doi: 10.1007/s13402-025-01149-3

Figure Lengend Snippet: VCP Dysfunction-induced mitochondrial aggregates initiated mitophagy. ( A ) U87 was treated with 10 μM CCCP, 15 μM V8, and 5 μM NMS873 for 3, 8, 12, 24 h, mitophagy-related proteins on mitochondria were quantified by WB. The indicated protein expression levels were quantified and normalized to COXIV. ( B - F ) The localization of PRKN ( B ), P62 ( C ), LC3 ( D ), BNIP3 ( E ), and TAXIBP1 ( F ) on mitochondria. U87 were transfected with pTagRFP-mito plasmid and then treated with 10 μM CCCP, 15 μM V8, and 5 μM NMS873 for 8 h. ( G ) The translocation of mitophagy-related proteins onto mitochondria. PRKN was knocked down by siRNA in U87 and then treated with 15 μM V8, or 5 μM NMS873 for 8 h. Mitochondrial proteins were extracted and analyzed by WB. The indicated protein expression levels were quantified and normalized to COXIV. ( H ) VCP overexpression inhibited the translocation of mitophagy-related proteins onto mitochondria. U87 and VCP overexpression U87 cells were treated with 15 μM V8, or 5 μM NMS873 for 8 h.The indicated protein expression levels were quantified and normalized to COXIV. The above mitochondrial protein samples derive from the same experiment. Bar, SD. * p < 0.05 or ** p < 0.01. “ns” means not significant (p > 0.05)

Article Snippet: Primary antibodies included those against ACTB/β-actin (ABclonal Technology, AC026), VCP(Proteintech, 10736–1-AP), XBP1S (Abcam, ab37152;1:2000), ATF4 (Proteintech, 10835–1-AP), P-EIF2α (Abclonal, AP0692), BiP/GRP78(Abclonal, A23453), IRE(Proteintech,27528–1-AP), K48-linkage Specific Ubiquitin (Abclonal, A3606), PINK1(Proteintech, 23274–1-AP), PRKN (Abways, CY6641), P62(Proteintech, 18420–1-AP),BNIP3(Abclonal, A5683), LC3(Proteintech, 14600–1-AP), COXIV (HUABIO, ET1701-63), TAXIBP1(Proteintech, 14424–1-AP), TUBLIN (Abclonal, A17545), Ubiquitin (Proteintech, 10201–2-AP).

Techniques: Expressing, Transfection, Plasmid Preparation, Translocation Assay, Over Expression

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: Targeting VCP with V8 suppresses glioblastoma development via formation of aggregates and disruption of mitophagy flux

doi: 10.1007/s13402-025-01149-3

Figure Lengend Snippet: V8 inhibited GBM growth in vivo. ( A - D ) The effects of V8 on U87 xenograft in vivo. Mice with U87 cell xenograft were treated with vehicle, V8, or TMZ for 14 days. Mice were sacrificed and tumor volumes ( B ), tumor weights ( C ) and body weight ( D ) were measured. N = 6, Two out of six mice in the TMZ-treated group achieved complete tumor regression (defined as tumor volume was non-detectable for 5 consecutive days). ( E and F ) Immunohistochemical staining documenting non-extractable VCP ( E ) and ubiquitinated proteins ( F ) in U87 xenografts from mice treated by V8. Mean density of VCP and UB was quantified by ImageJ. ( G - J ) Localization of VCP, PRKN, P62 and LC3 on mitochondria in tumor tissues. Tumor tissue was stained with LC3 ( G ), P62 ( I ), PRKN ( I ), VCP ( J ) and VDACI antibodies. Bar, SD. * p < 0.05 or ** p < 0.01. “ns” means not significant (p > 0.05). Data represent the mean ± SD. ( K ) Mice with U87 cell orthotopic xenograft were treated with vehicle and V8 for 14 days (N = 3). Tumor growth was monitored using in vivo bioluminescence imaging. ( L ) Representative images of KI67 staining in the brain tissue; control group (left) versus V8 group (right)

Article Snippet: Primary antibodies included those against ACTB/β-actin (ABclonal Technology, AC026), VCP(Proteintech, 10736–1-AP), XBP1S (Abcam, ab37152;1:2000), ATF4 (Proteintech, 10835–1-AP), P-EIF2α (Abclonal, AP0692), BiP/GRP78(Abclonal, A23453), IRE(Proteintech,27528–1-AP), K48-linkage Specific Ubiquitin (Abclonal, A3606), PINK1(Proteintech, 23274–1-AP), PRKN (Abways, CY6641), P62(Proteintech, 18420–1-AP),BNIP3(Abclonal, A5683), LC3(Proteintech, 14600–1-AP), COXIV (HUABIO, ET1701-63), TAXIBP1(Proteintech, 14424–1-AP), TUBLIN (Abclonal, A17545), Ubiquitin (Proteintech, 10201–2-AP).

Techniques: In Vivo, Immunohistochemical staining, Staining, Imaging, Control

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: Targeting VCP with V8 suppresses glioblastoma development via formation of aggregates and disruption of mitophagy flux

doi: 10.1007/s13402-025-01149-3

Figure Lengend Snippet: VCP Dysfunction-induced mitochondrial aggregates initiated mitophagy. ( A ) U87 was treated with 10 μM CCCP, 15 μM V8, and 5 μM NMS873 for 3, 8, 12, 24 h, mitophagy-related proteins on mitochondria were quantified by WB. The indicated protein expression levels were quantified and normalized to COXIV. ( B - F ) The localization of PRKN ( B ), P62 ( C ), LC3 ( D ), BNIP3 ( E ), and TAXIBP1 ( F ) on mitochondria. U87 were transfected with pTagRFP-mito plasmid and then treated with 10 μM CCCP, 15 μM V8, and 5 μM NMS873 for 8 h. ( G ) The translocation of mitophagy-related proteins onto mitochondria. PRKN was knocked down by siRNA in U87 and then treated with 15 μM V8, or 5 μM NMS873 for 8 h. Mitochondrial proteins were extracted and analyzed by WB. The indicated protein expression levels were quantified and normalized to COXIV. ( H ) VCP overexpression inhibited the translocation of mitophagy-related proteins onto mitochondria. U87 and VCP overexpression U87 cells were treated with 15 μM V8, or 5 μM NMS873 for 8 h.The indicated protein expression levels were quantified and normalized to COXIV. The above mitochondrial protein samples derive from the same experiment. Bar, SD. * p < 0.05 or ** p < 0.01. “ns” means not significant (p > 0.05)

Article Snippet: Primary antibodies included those against VCP (Proteintech, 10736–1-AP, 1:50) and PRKN (SELLECK, F0296; 1:50).

Techniques: Expressing, Transfection, Plasmid Preparation, Translocation Assay, Over Expression

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: Targeting VCP with V8 suppresses glioblastoma development via formation of aggregates and disruption of mitophagy flux

doi: 10.1007/s13402-025-01149-3

Figure Lengend Snippet: V8 inhibited GBM growth in vivo. ( A - D ) The effects of V8 on U87 xenograft in vivo. Mice with U87 cell xenograft were treated with vehicle, V8, or TMZ for 14 days. Mice were sacrificed and tumor volumes ( B ), tumor weights ( C ) and body weight ( D ) were measured. N = 6, Two out of six mice in the TMZ-treated group achieved complete tumor regression (defined as tumor volume was non-detectable for 5 consecutive days). ( E and F ) Immunohistochemical staining documenting non-extractable VCP ( E ) and ubiquitinated proteins ( F ) in U87 xenografts from mice treated by V8. Mean density of VCP and UB was quantified by ImageJ. ( G - J ) Localization of VCP, PRKN, P62 and LC3 on mitochondria in tumor tissues. Tumor tissue was stained with LC3 ( G ), P62 ( I ), PRKN ( I ), VCP ( J ) and VDACI antibodies. Bar, SD. * p < 0.05 or ** p < 0.01. “ns” means not significant (p > 0.05). Data represent the mean ± SD. ( K ) Mice with U87 cell orthotopic xenograft were treated with vehicle and V8 for 14 days (N = 3). Tumor growth was monitored using in vivo bioluminescence imaging. ( L ) Representative images of KI67 staining in the brain tissue; control group (left) versus V8 group (right)

Article Snippet: Primary antibodies included those against VCP (Proteintech, 10736–1-AP, 1:50) and PRKN (SELLECK, F0296; 1:50).

Techniques: In Vivo, Immunohistochemical staining, Staining, Imaging, Control