Journal: Biofactors (Oxford, England)

Article Title: ASNS Regulates H 2 O 2 ‐Induced Senescence, Oxidative Stress, and Glucose Metabolism in ARPE ‐19 Cells by Modulating USP13 Expression

doi: 10.1002/biof.70057

Figure Lengend Snippet: ASNS enhanced H 2 O 2 ‐induced retinal cell proliferation and attenuated senescence. ARPE‐19 cells were transfected with shASNS or ASNS‐OE. (A) The mRNA expression of ASNS was detected by RT‐qPCR. (B) The protein expression of ASNS was detected by WB. ARPE‐19 cells were divided into H 2 O 2 + shCtrl, H 2 O 2 + shASNS or H 2 O 2 + NC, H 2 O 2 + ASNS‐OE groups. (C) CCK8 assay was utilized to assess cell viability in H 2 O 2 ‐treated ARPE‐19 cells. (D) SA‐β‐gal staining experiments were conducted in the above groups. (E) Measurement of intracellular ROS in the above groups. (F) The protein expression of P53, P21, P16 was detected by WB. (G) The level of GSH and MDA detected by ELISA. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Lentiviral vectors interfering with ASNS and USP13 expression (shASNS and shUSP13) and the negative control shCtrl were synthesized by GeneChem (Shanghai, China).

Techniques: Transfection, Expressing, Quantitative RT-PCR, CCK-8 Assay, Staining, Enzyme-linked Immunosorbent Assay

Journal: Biofactors (Oxford, England)

Article Title: ASNS Regulates H 2 O 2 ‐Induced Senescence, Oxidative Stress, and Glucose Metabolism in ARPE ‐19 Cells by Modulating USP13 Expression

doi: 10.1002/biof.70057

Figure Lengend Snippet: ASNS regulated glucose metabolism pathways in H 2 O 2 ‐induced retinal cells. ARPE‐19 cells were divided into H 2 O 2 + shCtrl, H 2 O 2 + shASNS or H 2 O 2 + NC, H 2 O 2 + ASNS‐OE groups. (A) Detection of glucose uptake using a kit in H 2 O 2 ‐treated ARPE‐19 cells. (B) Detection of lactate production using a kit in H 2 O 2 ‐treated ARPE‐19 cells. (C) Measuring ECAR in H 2 O 2 ‐treated ARPE‐19 cells using the XF‐96 extracellular flux analyzer. (D) Measuring OCR in H 2 O 2 ‐treated ARPE‐19 cells using the XF‐96 extracellular flux analyzer. (E) The mRNA expression of Glut1, Glut4, HK2, and PGK1was detected by RT‐qPCR. (F) The protein expression of Glut1, Glut4, HK2, and PGK1was detected by WB. (G) The protein expression of HIF‐1α was detected by WB. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Lentiviral vectors interfering with ASNS and USP13 expression (shASNS and shUSP13) and the negative control shCtrl were synthesized by GeneChem (Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR

Journal: Biofactors (Oxford, England)

Article Title: ASNS Regulates H 2 O 2 ‐Induced Senescence, Oxidative Stress, and Glucose Metabolism in ARPE ‐19 Cells by Modulating USP13 Expression

doi: 10.1002/biof.70057

Figure Lengend Snippet: Validation of ASNS's impact on the AMD disease process in vivo. Sprague–Dawley rats injected with sodium iodate (30 mg/kg body weight) and divided into MOCK group (No lentivirus injections, n = 10), shCtrl group (Injection of shCtrl lentivirus, n = 10), and shASNS (Injection of shASNS lentivirus, n = 10) group. (A) H&E staining analysis of rat retinal tissues in each group. (B) SA‐β‐gal staining of rat retinal tissues were conducted in the above groups. (C) Measuring ECAR and OCR in retinal tissues using the XF‐96 extracellular flux analyzer. (D) The mRNA expression of Glut1, Glut4, HK2, and PGK1 in retinal tissues was detected by RT‐qPCR. (E) The protein expression of Glut1, Glut4, HK2, and PGK1 in retinal tissues was detected by WB. (F) The protein expression of ASNS and USP13 in retinal tissues was detected by WB. (G) IF was used to analyze the expression of ASNS and USP1 in retinal tissues. (H) The protein expression of HIF‐1α in retinal tissues was detected by WB. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Lentiviral vectors interfering with ASNS and USP13 expression (shASNS and shUSP13) and the negative control shCtrl were synthesized by GeneChem (Shanghai, China).

Techniques: Biomarker Discovery, In Vivo, Injection, Staining, Expressing, Quantitative RT-PCR

Journal: Biofactors (Oxford, England)

Article Title: ASNS Regulates H 2 O 2 ‐Induced Senescence, Oxidative Stress, and Glucose Metabolism in ARPE ‐19 Cells by Modulating USP13 Expression

doi: 10.1002/biof.70057

Figure Lengend Snippet: Validation of USP13's impact on the AMD disease process in vivo. Sprague–Dawley rats injected with sodium iodate (30 mg/kg body weigh) and divided into MOCK group (No lentivirus injections, n = 10), shCtrl group (Injection of shCtrl lentivirus, n = 10), and shUSP13 (Injection of shUSP13 lentivirus, n = 10) group. (A) H&E staining analysis of rat retinal tissues in each group. (B) SA‐β‐gal staining of rat retinal tissues were conducted in the above groups. (C) Measuring ECAR and OCR in retinal tissues using the XF‐96 extracellular flux analyzer. (D) The mRNA expression of Glut1, Glut4, HK2, and PGK1 in retinal tissues was detected by RT‐qPCR. (E) The protein expression of Glut1, Glut4, HK2, and PGK1 in retinal tissues was detected by WB. (F) The protein expression of USP13 in retinal tissues was detected by WB. (G) The protein expression of HIF‐1α in retinal tissues was detected by WB. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Lentiviral vectors interfering with ASNS and USP13 expression (shASNS and shUSP13) and the negative control shCtrl were synthesized by GeneChem (Shanghai, China).

Techniques: Biomarker Discovery, In Vivo, Injection, Staining, Expressing, Quantitative RT-PCR