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ANGPTL8 regulates the AMPK signaling pathway and interacts with <t>PirB</t> to modulate glucose metabolism and mitochondrial function. ( A , B ) Western blot analysis examines the impact of ANGPTL8 on the expression levels of AMPK signaling-related proteins in MC3T3-E1 cells and RAW 264.7 cells. ( C , D ) Co-IP assay detecting the interaction between PirB and ANGPTL8 in MC3T3-E1 cells and RAW 264.7 cells. Input represents the total protein <t>lysate;</t> <t>IgG</t> serves as the negative control. ( E , F ) Seahorse assay evaluates the influence of ANGPTL8 on glycolysis rates in cells after addition of AMPK activator. ( G , H ) Seahorse assay assesses the effect of ANGPTL8 on mitochondrial metabolic stress in cells after addition of AMPK activator. Data are presented as mean ± SD; ( N = 3). * p < 0.05, ** p < 0.01
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ANGPTL8 regulates the AMPK signaling pathway and interacts with <t>PirB</t> to modulate glucose metabolism and mitochondrial function. ( A , B ) Western blot analysis examines the impact of ANGPTL8 on the expression levels of AMPK signaling-related proteins in MC3T3-E1 cells and RAW 264.7 cells. ( C , D ) Co-IP assay detecting the interaction between PirB and ANGPTL8 in MC3T3-E1 cells and RAW 264.7 cells. Input represents the total protein <t>lysate;</t> <t>IgG</t> serves as the negative control. ( E , F ) Seahorse assay evaluates the influence of ANGPTL8 on glycolysis rates in cells after addition of AMPK activator. ( G , H ) Seahorse assay assesses the effect of ANGPTL8 on mitochondrial metabolic stress in cells after addition of AMPK activator. Data are presented as mean ± SD; ( N = 3). * p < 0.05, ** p < 0.01
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ANGPTL8 regulates the AMPK signaling pathway and interacts with <t>PirB</t> to modulate glucose metabolism and mitochondrial function. ( A , B ) Western blot analysis examines the impact of ANGPTL8 on the expression levels of AMPK signaling-related proteins in MC3T3-E1 cells and RAW 264.7 cells. ( C , D ) Co-IP assay detecting the interaction between PirB and ANGPTL8 in MC3T3-E1 cells and RAW 264.7 cells. Input represents the total protein <t>lysate;</t> <t>IgG</t> serves as the negative control. ( E , F ) Seahorse assay evaluates the influence of ANGPTL8 on glycolysis rates in cells after addition of AMPK activator. ( G , H ) Seahorse assay assesses the effect of ANGPTL8 on mitochondrial metabolic stress in cells after addition of AMPK activator. Data are presented as mean ± SD; ( N = 3). * p < 0.05, ** p < 0.01
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ImmunoWay Biotechnology Company anti-pirb antibody
ANGPTL8 regulates the AMPK signaling pathway and interacts with <t>PirB</t> to modulate glucose metabolism and mitochondrial function. ( A , B ) Western blot analysis examines the impact of ANGPTL8 on the expression levels of AMPK signaling-related proteins in MC3T3-E1 cells and RAW 264.7 cells. ( C , D ) Co-IP assay detecting the interaction between PirB and ANGPTL8 in MC3T3-E1 cells and RAW 264.7 cells. Input represents the total protein <t>lysate;</t> <t>IgG</t> serves as the negative control. ( E , F ) Seahorse assay evaluates the influence of ANGPTL8 on glycolysis rates in cells after addition of AMPK activator. ( G , H ) Seahorse assay assesses the effect of ANGPTL8 on mitochondrial metabolic stress in cells after addition of AMPK activator. Data are presented as mean ± SD; ( N = 3). * p < 0.05, ** p < 0.01
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ANGPTL8 regulates the AMPK signaling pathway and interacts with PirB to modulate glucose metabolism and mitochondrial function. ( A , B ) Western blot analysis examines the impact of ANGPTL8 on the expression levels of AMPK signaling-related proteins in MC3T3-E1 cells and RAW 264.7 cells. ( C , D ) Co-IP assay detecting the interaction between PirB and ANGPTL8 in MC3T3-E1 cells and RAW 264.7 cells. Input represents the total protein lysate; IgG serves as the negative control. ( E , F ) Seahorse assay evaluates the influence of ANGPTL8 on glycolysis rates in cells after addition of AMPK activator. ( G , H ) Seahorse assay assesses the effect of ANGPTL8 on mitochondrial metabolic stress in cells after addition of AMPK activator. Data are presented as mean ± SD; ( N = 3). * p < 0.05, ** p < 0.01

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: ANGPTL8 accelerates bone loss in diabetic mice by promoting osteoclastic differentiation and inhibiting osteoblastic differentiation through AMPK pathway-mediated metabolic reprogramming

doi: 10.1007/s00018-025-06077-x

Figure Lengend Snippet: ANGPTL8 regulates the AMPK signaling pathway and interacts with PirB to modulate glucose metabolism and mitochondrial function. ( A , B ) Western blot analysis examines the impact of ANGPTL8 on the expression levels of AMPK signaling-related proteins in MC3T3-E1 cells and RAW 264.7 cells. ( C , D ) Co-IP assay detecting the interaction between PirB and ANGPTL8 in MC3T3-E1 cells and RAW 264.7 cells. Input represents the total protein lysate; IgG serves as the negative control. ( E , F ) Seahorse assay evaluates the influence of ANGPTL8 on glycolysis rates in cells after addition of AMPK activator. ( G , H ) Seahorse assay assesses the effect of ANGPTL8 on mitochondrial metabolic stress in cells after addition of AMPK activator. Data are presented as mean ± SD; ( N = 3). * p < 0.05, ** p < 0.01

Article Snippet: The supernatant was incubated overnight at 4 °C with either control IgG antibody or primary antibodies against ANGPTL8 (1:1000 dilution; Proteintech, 23792-1-AP) or PirB.

Techniques: Western Blot, Expressing, Co-Immunoprecipitation Assay, Negative Control

PirB knockdown modulates ANGPTL8-mediated functional and metabolic phenotypes in MC3T3-E1 cells and RAW 264.7 cells by regulating the AMPK signaling pathway. ( A , B ) Western blot analysis the impact of si-PirB on the expression levels of AMPK signaling-related proteins in MC3T3-E1 cells and RAW 264.7 cells. (C, D) RT-qPCR analysis the impact of si-PirB on the mRNA levels in differentiated cells treated wtih high glucose and ANGPTL8 protein. ( E ) The impact of PirB knockdown on lactate levels in ANGPTL8 protein-treated cells with AMPK activator. ( F ) The effects of PirB knockdown on GSH levels in ANGPTL8 protein-treated cells with AMPK activator. ( G ) The effects of PirB knockdown on the ratio of NAD + /NADH in ANGPTL8 protein-treated cells with AMPK activator. (H) The effects of PirB knockdown on ROS levels in ANGPTL8 protein-treated cells with AMPK activator. Data are presented as mean ± SD; ( N = 3). ns: not significant, * p < 0.05, ** p < 0.01

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: ANGPTL8 accelerates bone loss in diabetic mice by promoting osteoclastic differentiation and inhibiting osteoblastic differentiation through AMPK pathway-mediated metabolic reprogramming

doi: 10.1007/s00018-025-06077-x

Figure Lengend Snippet: PirB knockdown modulates ANGPTL8-mediated functional and metabolic phenotypes in MC3T3-E1 cells and RAW 264.7 cells by regulating the AMPK signaling pathway. ( A , B ) Western blot analysis the impact of si-PirB on the expression levels of AMPK signaling-related proteins in MC3T3-E1 cells and RAW 264.7 cells. (C, D) RT-qPCR analysis the impact of si-PirB on the mRNA levels in differentiated cells treated wtih high glucose and ANGPTL8 protein. ( E ) The impact of PirB knockdown on lactate levels in ANGPTL8 protein-treated cells with AMPK activator. ( F ) The effects of PirB knockdown on GSH levels in ANGPTL8 protein-treated cells with AMPK activator. ( G ) The effects of PirB knockdown on the ratio of NAD + /NADH in ANGPTL8 protein-treated cells with AMPK activator. (H) The effects of PirB knockdown on ROS levels in ANGPTL8 protein-treated cells with AMPK activator. Data are presented as mean ± SD; ( N = 3). ns: not significant, * p < 0.05, ** p < 0.01

Article Snippet: The supernatant was incubated overnight at 4 °C with either control IgG antibody or primary antibodies against ANGPTL8 (1:1000 dilution; Proteintech, 23792-1-AP) or PirB.

Techniques: Knockdown, Functional Assay, Western Blot, Expressing, Quantitative RT-PCR