Journal: BMC Cancer

Article Title: ATP6V0B promotes the tumorigenesis of bladder cancer by activating PAQR4/PI3K/AKT signaling

doi: 10.1186/s12885-025-14183-z

Figure Lengend Snippet: ATP6V0B was upregulated in BCa. The GEPIA database was used to determine the mRNA level of ATP6V0B ( A ) and the overall survival of BCa patients ( B ). ( C ) The Kaplan-Meier (K-M) plotter was used to analyze the mRNA level of ATP6V0B and survival data. ( D ) RT-qPCR assays were performed to determine the mRNA level of ATP6V0B in BCa tumor tissues and adjacent normal tissues ( n = 30). ( E ) Western blotting assays were conducted to quantify the expression of ATP6V0B in four paired BCa tumor tissues and adjacent normal tissues. ( F ) Western blotting assays were performed to quantify the expression of ATP6V0B in a non-cancerous immortalized urothelial cell line SV-Huc-1 and four BCa cell lines, including J82, T24, UMUC3, and 5637; * p < 0.05 and ** p < 0.01

Article Snippet: The primary antibodies used in this study are as follows: ATP6V0B rabbit antibody (Cat No: YN3416, 1:1000, Immunoway, China), β-Tubulin polyclonal antibody (Cat No: AC015, 1:5000, ABclonal, China), p85 monoclonal antibody (Cat No: A11177, 1:1000, ABclonal, China), p-p85 antibody (Cat No: ab191606, 1:1000, Abcam, USA), AKT antibody (Cat No: ab8805, 1:500, Abcam, USA), p-AKT (Ser473) antibody (Cat No: 80455-1-RR, 1:5000, Proteintech, China), E-cadherin antibody (Cat No: EPR16845-108, 1:1000, Abcam, USA), Vimentin (Cat No: A11952, 1:1000, ABclonal, China), and PAQR4 (Cat No: SAB2107001, 1:1000, Sigma-Aldrich, USA).

Techniques: Quantitative RT-PCR, Western Blot, Expressing

Journal: BMC Cancer

Article Title: ATP6V0B promotes the tumorigenesis of bladder cancer by activating PAQR4/PI3K/AKT signaling

doi: 10.1186/s12885-025-14183-z

Figure Lengend Snippet: ATP6V0B increased the proliferation, migration, and invasion abilities of BCa cells in vitro. ( A ) The efficiency of the overexpression and knockdown of ATP6V0B plasmids was evaluated by RT-qPCR assays in BCa cells. ( B - D ) The CCK-8 assay was performed to measure the effects of the overexpression and knockdown of ATP6V0B on the proliferation of BCa cells. ( E ) The migration and invasion abilities of BCa cells were evaluated by overexpressing and knocking down ATP6V0B. ( F ) Western blotting analysis was performed to quantify the expression of ATP6V0B, E-cadherin, and Vimentin. ( G - I ) Quantification of the protein levels in ( F ); * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: The primary antibodies used in this study are as follows: ATP6V0B rabbit antibody (Cat No: YN3416, 1:1000, Immunoway, China), β-Tubulin polyclonal antibody (Cat No: AC015, 1:5000, ABclonal, China), p85 monoclonal antibody (Cat No: A11177, 1:1000, ABclonal, China), p-p85 antibody (Cat No: ab191606, 1:1000, Abcam, USA), AKT antibody (Cat No: ab8805, 1:500, Abcam, USA), p-AKT (Ser473) antibody (Cat No: 80455-1-RR, 1:5000, Proteintech, China), E-cadherin antibody (Cat No: EPR16845-108, 1:1000, Abcam, USA), Vimentin (Cat No: A11952, 1:1000, ABclonal, China), and PAQR4 (Cat No: SAB2107001, 1:1000, Sigma-Aldrich, USA).

Techniques: Migration, In Vitro, Over Expression, Knockdown, Quantitative RT-PCR, CCK-8 Assay, Western Blot, Expressing

Journal: BMC Cancer

Article Title: ATP6V0B promotes the tumorigenesis of bladder cancer by activating PAQR4/PI3K/AKT signaling

doi: 10.1186/s12885-025-14183-z

Figure Lengend Snippet: Knocking down ATP6V0B suppressed tumor growth in vivo. ( A ) The representative photographs of tumors were taken from nude mice 21 days after inoculation ( n = 6). ( B and C ) The volume and weight of the tumors were measured after the tumors were isolated. ( D ) RT-qPCR analysis was performed to evaluate the mRNA levels of ATP6V0B in tumor tissues. ( E ) The expression of ATP6V0B in tumor tissues was confirmed by IHC; **** p < 0.0001

Article Snippet: The primary antibodies used in this study are as follows: ATP6V0B rabbit antibody (Cat No: YN3416, 1:1000, Immunoway, China), β-Tubulin polyclonal antibody (Cat No: AC015, 1:5000, ABclonal, China), p85 monoclonal antibody (Cat No: A11177, 1:1000, ABclonal, China), p-p85 antibody (Cat No: ab191606, 1:1000, Abcam, USA), AKT antibody (Cat No: ab8805, 1:500, Abcam, USA), p-AKT (Ser473) antibody (Cat No: 80455-1-RR, 1:5000, Proteintech, China), E-cadherin antibody (Cat No: EPR16845-108, 1:1000, Abcam, USA), Vimentin (Cat No: A11952, 1:1000, ABclonal, China), and PAQR4 (Cat No: SAB2107001, 1:1000, Sigma-Aldrich, USA).

Techniques: In Vivo, Isolation, Quantitative RT-PCR, Expressing, Paraffin-embedded Immunohistochemistry

Journal: BMC Cancer

Article Title: ATP6V0B promotes the tumorigenesis of bladder cancer by activating PAQR4/PI3K/AKT signaling

doi: 10.1186/s12885-025-14183-z

Figure Lengend Snippet: ATP6V0B activated PI3K/AKT signaling in BCa cells. ( A ) GSEA results showed that the PI3K/AKT signaling pathway was hyperactivated in BCa cells. ( B ) Western blotting assays were performed to determine the indicated protein levels of PI3K/AKT signaling. ( C ) Quantification of the protein expression levels in ( B ). ( D ) The BCa cells were transfected with the pcDNA3 empty vector and the ATP6V0B-overexpressing vector for 24 h, after which the cells were treated with LY294002 (10 µM) for 24 h. Western blotting assays were performed to determine the level of p85, p-p85, AKT, and p-AKT proteins. ( E ) Quantification of the protein levels in ( D ); * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; ns, no significant difference

Article Snippet: The primary antibodies used in this study are as follows: ATP6V0B rabbit antibody (Cat No: YN3416, 1:1000, Immunoway, China), β-Tubulin polyclonal antibody (Cat No: AC015, 1:5000, ABclonal, China), p85 monoclonal antibody (Cat No: A11177, 1:1000, ABclonal, China), p-p85 antibody (Cat No: ab191606, 1:1000, Abcam, USA), AKT antibody (Cat No: ab8805, 1:500, Abcam, USA), p-AKT (Ser473) antibody (Cat No: 80455-1-RR, 1:5000, Proteintech, China), E-cadherin antibody (Cat No: EPR16845-108, 1:1000, Abcam, USA), Vimentin (Cat No: A11952, 1:1000, ABclonal, China), and PAQR4 (Cat No: SAB2107001, 1:1000, Sigma-Aldrich, USA).

Techniques: Western Blot, Expressing, Transfection, Plasmid Preparation

Journal: BMC Cancer

Article Title: ATP6V0B promotes the tumorigenesis of bladder cancer by activating PAQR4/PI3K/AKT signaling

doi: 10.1186/s12885-025-14183-z

Figure Lengend Snippet: ATP6V0B upregulated the expression of PAQR4 in BCa. ( A ) The Venn diagram shows that there are 54 genes that are positively correlated with ATP6V0B and upregulated in BCa, and among these 54 genes, 15 genes have significantly different survival curves. ( B - D ) The GEPIA database was used to analyze the correlation between ATP6V0B and PAQR4 ( B ), the level of PAQR4 mRNA ( C ), and overall survival, as determined by the level of PAQR4 mRNA ( D ). ( E ) The K-M plotter was used to validate the overall survival of PAQR4. ( F ) RT-qPCR analysis was performed to evaluate the level of PAQR4 mRNA in BCa tumor tissues and adjacent normal tissues ( n = 30). ( G ) The correlation between ATP6V0B and PAQR4 was analyzed based on the level of mRNA of ATP6V0B and PAQR4. ( H ) BCa cells were transfected with the indicated plasmids, and RT-qPCR analysis was performed to evaluate the level of PAQR4 mRNA 36 h after transfection. ( I ) Western blotting analysis was performed to measure the expression of PAQR4 48 h after transfection; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: The primary antibodies used in this study are as follows: ATP6V0B rabbit antibody (Cat No: YN3416, 1:1000, Immunoway, China), β-Tubulin polyclonal antibody (Cat No: AC015, 1:5000, ABclonal, China), p85 monoclonal antibody (Cat No: A11177, 1:1000, ABclonal, China), p-p85 antibody (Cat No: ab191606, 1:1000, Abcam, USA), AKT antibody (Cat No: ab8805, 1:500, Abcam, USA), p-AKT (Ser473) antibody (Cat No: 80455-1-RR, 1:5000, Proteintech, China), E-cadherin antibody (Cat No: EPR16845-108, 1:1000, Abcam, USA), Vimentin (Cat No: A11952, 1:1000, ABclonal, China), and PAQR4 (Cat No: SAB2107001, 1:1000, Sigma-Aldrich, USA).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot

Journal: BMC Cancer

Article Title: ATP6V0B promotes the tumorigenesis of bladder cancer by activating PAQR4/PI3K/AKT signaling

doi: 10.1186/s12885-025-14183-z

Figure Lengend Snippet: ATP6V0B increased the malignant behavior of BCa cells and activation of PI3K/AKT signaling by upregulating PAQR4. BCa cells were transfected with pcDNA3 + pSilencer2.1, ATP6V0B + pSilencer2.1, and ATP6V0B + pshR-PAQR4, after which the abilities of the cells to proliferate ( A ), migrate, and invade ( B ) were evaluated. ( C ) Western blotting assays were conducted to determine the levels of the proteins associated with the PI3K/AKT signaling pathway affected by the overexpression of PAQR4. ( D ) Quantification of the protein levels in ( C ); * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; ns, no significant difference

Article Snippet: The primary antibodies used in this study are as follows: ATP6V0B rabbit antibody (Cat No: YN3416, 1:1000, Immunoway, China), β-Tubulin polyclonal antibody (Cat No: AC015, 1:5000, ABclonal, China), p85 monoclonal antibody (Cat No: A11177, 1:1000, ABclonal, China), p-p85 antibody (Cat No: ab191606, 1:1000, Abcam, USA), AKT antibody (Cat No: ab8805, 1:500, Abcam, USA), p-AKT (Ser473) antibody (Cat No: 80455-1-RR, 1:5000, Proteintech, China), E-cadherin antibody (Cat No: EPR16845-108, 1:1000, Abcam, USA), Vimentin (Cat No: A11952, 1:1000, ABclonal, China), and PAQR4 (Cat No: SAB2107001, 1:1000, Sigma-Aldrich, USA).

Techniques: Activation Assay, Transfection, Western Blot, Over Expression

Journal: BMC Cancer

Article Title: ATP6V0B promotes the tumorigenesis of bladder cancer by activating PAQR4/PI3K/AKT signaling

doi: 10.1186/s12885-025-14183-z

Figure Lengend Snippet: ATP6V0B was upregulated in BCa. The GEPIA database was used to determine the mRNA level of ATP6V0B ( A ) and the overall survival of BCa patients ( B ). ( C ) The Kaplan-Meier (K-M) plotter was used to analyze the mRNA level of ATP6V0B and survival data. ( D ) RT-qPCR assays were performed to determine the mRNA level of ATP6V0B in BCa tumor tissues and adjacent normal tissues ( n = 30). ( E ) Western blotting assays were conducted to quantify the expression of ATP6V0B in four paired BCa tumor tissues and adjacent normal tissues. ( F ) Western blotting assays were performed to quantify the expression of ATP6V0B in a non-cancerous immortalized urothelial cell line SV-Huc-1 and four BCa cell lines, including J82, T24, UMUC3, and 5637; * p < 0.05 and ** p < 0.01

Article Snippet: Subsequently, the sections were blocked with a 5% BSA and incubated at 4 °C overnight with rabbit anti-ATP6V0B antibody (Cat No: YN3416, 1:100, Immunoway, China) and Ki67Ki67 antibody (Cat No: ab15580, 1:500, Abcam, USA).

Techniques: Quantitative RT-PCR, Western Blot, Expressing

Journal: BMC Cancer

Article Title: ATP6V0B promotes the tumorigenesis of bladder cancer by activating PAQR4/PI3K/AKT signaling

doi: 10.1186/s12885-025-14183-z

Figure Lengend Snippet: ATP6V0B increased the proliferation, migration, and invasion abilities of BCa cells in vitro. ( A ) The efficiency of the overexpression and knockdown of ATP6V0B plasmids was evaluated by RT-qPCR assays in BCa cells. ( B - D ) The CCK-8 assay was performed to measure the effects of the overexpression and knockdown of ATP6V0B on the proliferation of BCa cells. ( E ) The migration and invasion abilities of BCa cells were evaluated by overexpressing and knocking down ATP6V0B. ( F ) Western blotting analysis was performed to quantify the expression of ATP6V0B, E-cadherin, and Vimentin. ( G - I ) Quantification of the protein levels in ( F ); * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: Subsequently, the sections were blocked with a 5% BSA and incubated at 4 °C overnight with rabbit anti-ATP6V0B antibody (Cat No: YN3416, 1:100, Immunoway, China) and Ki67Ki67 antibody (Cat No: ab15580, 1:500, Abcam, USA).

Techniques: Migration, In Vitro, Over Expression, Knockdown, Quantitative RT-PCR, CCK-8 Assay, Western Blot, Expressing

Journal: BMC Cancer

Article Title: ATP6V0B promotes the tumorigenesis of bladder cancer by activating PAQR4/PI3K/AKT signaling

doi: 10.1186/s12885-025-14183-z

Figure Lengend Snippet: Knocking down ATP6V0B suppressed tumor growth in vivo. ( A ) The representative photographs of tumors were taken from nude mice 21 days after inoculation ( n = 6). ( B and C ) The volume and weight of the tumors were measured after the tumors were isolated. ( D ) RT-qPCR analysis was performed to evaluate the mRNA levels of ATP6V0B in tumor tissues. ( E ) The expression of ATP6V0B in tumor tissues was confirmed by IHC; **** p < 0.0001

Article Snippet: Subsequently, the sections were blocked with a 5% BSA and incubated at 4 °C overnight with rabbit anti-ATP6V0B antibody (Cat No: YN3416, 1:100, Immunoway, China) and Ki67Ki67 antibody (Cat No: ab15580, 1:500, Abcam, USA).

Techniques: In Vivo, Isolation, Quantitative RT-PCR, Expressing, Paraffin-embedded Immunohistochemistry

Journal: BMC Cancer

Article Title: ATP6V0B promotes the tumorigenesis of bladder cancer by activating PAQR4/PI3K/AKT signaling

doi: 10.1186/s12885-025-14183-z

Figure Lengend Snippet: ATP6V0B activated PI3K/AKT signaling in BCa cells. ( A ) GSEA results showed that the PI3K/AKT signaling pathway was hyperactivated in BCa cells. ( B ) Western blotting assays were performed to determine the indicated protein levels of PI3K/AKT signaling. ( C ) Quantification of the protein expression levels in ( B ). ( D ) The BCa cells were transfected with the pcDNA3 empty vector and the ATP6V0B-overexpressing vector for 24 h, after which the cells were treated with LY294002 (10 µM) for 24 h. Western blotting assays were performed to determine the level of p85, p-p85, AKT, and p-AKT proteins. ( E ) Quantification of the protein levels in ( D ); * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; ns, no significant difference

Article Snippet: Subsequently, the sections were blocked with a 5% BSA and incubated at 4 °C overnight with rabbit anti-ATP6V0B antibody (Cat No: YN3416, 1:100, Immunoway, China) and Ki67Ki67 antibody (Cat No: ab15580, 1:500, Abcam, USA).

Techniques: Western Blot, Expressing, Transfection, Plasmid Preparation

Journal: BMC Cancer

Article Title: ATP6V0B promotes the tumorigenesis of bladder cancer by activating PAQR4/PI3K/AKT signaling

doi: 10.1186/s12885-025-14183-z

Figure Lengend Snippet: ATP6V0B upregulated the expression of PAQR4 in BCa. ( A ) The Venn diagram shows that there are 54 genes that are positively correlated with ATP6V0B and upregulated in BCa, and among these 54 genes, 15 genes have significantly different survival curves. ( B - D ) The GEPIA database was used to analyze the correlation between ATP6V0B and PAQR4 ( B ), the level of PAQR4 mRNA ( C ), and overall survival, as determined by the level of PAQR4 mRNA ( D ). ( E ) The K-M plotter was used to validate the overall survival of PAQR4. ( F ) RT-qPCR analysis was performed to evaluate the level of PAQR4 mRNA in BCa tumor tissues and adjacent normal tissues ( n = 30). ( G ) The correlation between ATP6V0B and PAQR4 was analyzed based on the level of mRNA of ATP6V0B and PAQR4. ( H ) BCa cells were transfected with the indicated plasmids, and RT-qPCR analysis was performed to evaluate the level of PAQR4 mRNA 36 h after transfection. ( I ) Western blotting analysis was performed to measure the expression of PAQR4 48 h after transfection; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: Subsequently, the sections were blocked with a 5% BSA and incubated at 4 °C overnight with rabbit anti-ATP6V0B antibody (Cat No: YN3416, 1:100, Immunoway, China) and Ki67Ki67 antibody (Cat No: ab15580, 1:500, Abcam, USA).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot

Journal: BMC Cancer

Article Title: ATP6V0B promotes the tumorigenesis of bladder cancer by activating PAQR4/PI3K/AKT signaling

doi: 10.1186/s12885-025-14183-z

Figure Lengend Snippet: ATP6V0B increased the malignant behavior of BCa cells and activation of PI3K/AKT signaling by upregulating PAQR4. BCa cells were transfected with pcDNA3 + pSilencer2.1, ATP6V0B + pSilencer2.1, and ATP6V0B + pshR-PAQR4, after which the abilities of the cells to proliferate ( A ), migrate, and invade ( B ) were evaluated. ( C ) Western blotting assays were conducted to determine the levels of the proteins associated with the PI3K/AKT signaling pathway affected by the overexpression of PAQR4. ( D ) Quantification of the protein levels in ( C ); * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; ns, no significant difference

Article Snippet: Subsequently, the sections were blocked with a 5% BSA and incubated at 4 °C overnight with rabbit anti-ATP6V0B antibody (Cat No: YN3416, 1:100, Immunoway, China) and Ki67Ki67 antibody (Cat No: ab15580, 1:500, Abcam, USA).

Techniques: Activation Assay, Transfection, Western Blot, Over Expression

Journal: Oncology Letters

Article Title: Dihydroartemisinin inhibits ATP6 activity, reduces energy metabolism of hepatocellular carcinoma cells, promotes apoptosis and inhibits metastasis via CANX

doi: 10.3892/ol.2024.14607

Figure Lengend Snippet: Effects of CANX and DHA on ATP6V0B and ATP6V1F expression. (A) Expression of (B) CANX, (C) ATP6V0B and (D) ATP6V1F in HuH-7 cells, detected by western blotting after CANX knockout, DHA or CANX overexpression. (E) Expression of (F) CANX, (G) ATP6V0B and (H) ATP6V1F in Li-7 cells, detected by western blotting after CANX knockout, DHA or CANX overexpression. **P<0.01; ***P<0.001. CANX, calnexin; DHA, dihydroartemisinin; NC, negative control; siRNA, small interfering RNA.

Article Snippet: The primary antibodies used were CANX (1:5,000; cat. no. 66903-1-Ig; Proteintech Group, Inc.), ATP6V0B (1:1,000; cat. no. 27671-1-AP; Proteintech Group, Inc.) and ATP6V1F (1:1,000; ab190789; Abcam), β-actin (1:2,000; cat. no. 20536-1-AP; Proteintech Group, Inc.).

Techniques: Expressing, Western Blot, Knock-Out, Over Expression, Negative Control, Small Interfering RNA