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The mitophagy-driven metabolic–epigenetic axis suppresses SASP factors in senescent NP cells. (A) Western blot analysis of SASP factors and <t>thrombospondin-1</t> <t>(THBS1)</t> in young and aged NP cells. (B) Schematic illustration of the experimental design. (C) Western blot analysis of SASP factors and THBS1 in aged NP cells treated with vehicle, TJ0113 (5 μM), FX-11(20 μM), or both. (D) Western blot analysis of SASP factors and THBS1 in aged NP cells treated with vehicle, TJ0113 (5 <t>μM),</t> <t>L-Lac</t> (10 μM), or both. (E) qRT-PCR analysis of SASP factors in aged NP cells with treatment as (B) and (C) ( n = 3). (F) Western blot analysis of SASP factors and THBS1 in control and THBS1 knockdown aged NP cells treated with or without TJ0113 (5 μM, 48 h). (G) qRT-PCR analysis of SASP factors in control and THBS1 knockdown aged NP cells treated with TJ0113 or not (5 μM, 48 h) ( n = 3). (H) Western blot analysis of SASP factors and THBS1 in control and THBS1-treated (0.1/1 μg/ml, 48 h) aged NP cells under treatment with or without TJ0113 (5 μM, 48 h). (I) qRT-PCR analysis of SASP factors in control and THBS1-treated (0.1/1 μg/ml, 48 h) aged NP cells under treatment with or without TJ0113 (5 μM, 48 h) ( n = 3). Results represent the mean ± SD of at least 3 independent experiments. Significance levels are shown within the graphs.
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The mitophagy-driven metabolic–epigenetic axis suppresses SASP factors in senescent NP cells. (A) Western blot analysis of SASP factors and <t>thrombospondin-1</t> <t>(THBS1)</t> in young and aged NP cells. (B) Schematic illustration of the experimental design. (C) Western blot analysis of SASP factors and THBS1 in aged NP cells treated with vehicle, TJ0113 (5 μM), FX-11(20 μM), or both. (D) Western blot analysis of SASP factors and THBS1 in aged NP cells treated with vehicle, TJ0113 (5 <t>μM),</t> <t>L-Lac</t> (10 μM), or both. (E) qRT-PCR analysis of SASP factors in aged NP cells with treatment as (B) and (C) ( n = 3). (F) Western blot analysis of SASP factors and THBS1 in control and THBS1 knockdown aged NP cells treated with or without TJ0113 (5 μM, 48 h). (G) qRT-PCR analysis of SASP factors in control and THBS1 knockdown aged NP cells treated with TJ0113 or not (5 μM, 48 h) ( n = 3). (H) Western blot analysis of SASP factors and THBS1 in control and THBS1-treated (0.1/1 μg/ml, 48 h) aged NP cells under treatment with or without TJ0113 (5 μM, 48 h). (I) qRT-PCR analysis of SASP factors in control and THBS1-treated (0.1/1 μg/ml, 48 h) aged NP cells under treatment with or without TJ0113 (5 μM, 48 h) ( n = 3). Results represent the mean ± SD of at least 3 independent experiments. Significance levels are shown within the graphs.
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The mitophagy-driven metabolic–epigenetic axis suppresses SASP factors in senescent NP cells. (A) Western blot analysis of SASP factors and <t>thrombospondin-1</t> <t>(THBS1)</t> in young and aged NP cells. (B) Schematic illustration of the experimental design. (C) Western blot analysis of SASP factors and THBS1 in aged NP cells treated with vehicle, TJ0113 (5 μM), FX-11(20 μM), or both. (D) Western blot analysis of SASP factors and THBS1 in aged NP cells treated with vehicle, TJ0113 (5 <t>μM),</t> <t>L-Lac</t> (10 μM), or both. (E) qRT-PCR analysis of SASP factors in aged NP cells with treatment as (B) and (C) ( n = 3). (F) Western blot analysis of SASP factors and THBS1 in control and THBS1 knockdown aged NP cells treated with or without TJ0113 (5 μM, 48 h). (G) qRT-PCR analysis of SASP factors in control and THBS1 knockdown aged NP cells treated with TJ0113 or not (5 μM, 48 h) ( n = 3). (H) Western blot analysis of SASP factors and THBS1 in control and THBS1-treated (0.1/1 μg/ml, 48 h) aged NP cells under treatment with or without TJ0113 (5 μM, 48 h). (I) qRT-PCR analysis of SASP factors in control and THBS1-treated (0.1/1 μg/ml, 48 h) aged NP cells under treatment with or without TJ0113 (5 μM, 48 h) ( n = 3). Results represent the mean ± SD of at least 3 independent experiments. Significance levels are shown within the graphs.
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The mitophagy-driven metabolic–epigenetic axis suppresses SASP factors in senescent NP cells. (A) Western blot analysis of SASP factors and <t>thrombospondin-1</t> <t>(THBS1)</t> in young and aged NP cells. (B) Schematic illustration of the experimental design. (C) Western blot analysis of SASP factors and THBS1 in aged NP cells treated with vehicle, TJ0113 (5 μM), FX-11(20 μM), or both. (D) Western blot analysis of SASP factors and THBS1 in aged NP cells treated with vehicle, TJ0113 (5 <t>μM),</t> <t>L-Lac</t> (10 μM), or both. (E) qRT-PCR analysis of SASP factors in aged NP cells with treatment as (B) and (C) ( n = 3). (F) Western blot analysis of SASP factors and THBS1 in control and THBS1 knockdown aged NP cells treated with or without TJ0113 (5 μM, 48 h). (G) qRT-PCR analysis of SASP factors in control and THBS1 knockdown aged NP cells treated with TJ0113 or not (5 μM, 48 h) ( n = 3). (H) Western blot analysis of SASP factors and THBS1 in control and THBS1-treated (0.1/1 μg/ml, 48 h) aged NP cells under treatment with or without TJ0113 (5 μM, 48 h). (I) qRT-PCR analysis of SASP factors in control and THBS1-treated (0.1/1 μg/ml, 48 h) aged NP cells under treatment with or without TJ0113 (5 μM, 48 h) ( n = 3). Results represent the mean ± SD of at least 3 independent experiments. Significance levels are shown within the graphs.
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The mitophagy-driven metabolic–epigenetic axis suppresses SASP factors in senescent NP cells. (A) Western blot analysis of SASP factors and <t>thrombospondin-1</t> <t>(THBS1)</t> in young and aged NP cells. (B) Schematic illustration of the experimental design. (C) Western blot analysis of SASP factors and THBS1 in aged NP cells treated with vehicle, TJ0113 (5 μM), FX-11(20 μM), or both. (D) Western blot analysis of SASP factors and THBS1 in aged NP cells treated with vehicle, TJ0113 (5 <t>μM),</t> <t>L-Lac</t> (10 μM), or both. (E) qRT-PCR analysis of SASP factors in aged NP cells with treatment as (B) and (C) ( n = 3). (F) Western blot analysis of SASP factors and THBS1 in control and THBS1 knockdown aged NP cells treated with or without TJ0113 (5 μM, 48 h). (G) qRT-PCR analysis of SASP factors in control and THBS1 knockdown aged NP cells treated with TJ0113 or not (5 μM, 48 h) ( n = 3). (H) Western blot analysis of SASP factors and THBS1 in control and THBS1-treated (0.1/1 μg/ml, 48 h) aged NP cells under treatment with or without TJ0113 (5 μM, 48 h). (I) qRT-PCR analysis of SASP factors in control and THBS1-treated (0.1/1 μg/ml, 48 h) aged NP cells under treatment with or without TJ0113 (5 μM, 48 h) ( n = 3). Results represent the mean ± SD of at least 3 independent experiments. Significance levels are shown within the graphs.
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The mitophagy-driven metabolic–epigenetic axis suppresses SASP factors in senescent NP cells. (A) Western blot analysis of SASP factors and <t>thrombospondin-1</t> <t>(THBS1)</t> in young and aged NP cells. (B) Schematic illustration of the experimental design. (C) Western blot analysis of SASP factors and THBS1 in aged NP cells treated with vehicle, TJ0113 (5 μM), FX-11(20 μM), or both. (D) Western blot analysis of SASP factors and THBS1 in aged NP cells treated with vehicle, TJ0113 (5 <t>μM),</t> <t>L-Lac</t> (10 μM), or both. (E) qRT-PCR analysis of SASP factors in aged NP cells with treatment as (B) and (C) ( n = 3). (F) Western blot analysis of SASP factors and THBS1 in control and THBS1 knockdown aged NP cells treated with or without TJ0113 (5 μM, 48 h). (G) qRT-PCR analysis of SASP factors in control and THBS1 knockdown aged NP cells treated with TJ0113 or not (5 μM, 48 h) ( n = 3). (H) Western blot analysis of SASP factors and THBS1 in control and THBS1-treated (0.1/1 μg/ml, 48 h) aged NP cells under treatment with or without TJ0113 (5 μM, 48 h). (I) qRT-PCR analysis of SASP factors in control and THBS1-treated (0.1/1 μg/ml, 48 h) aged NP cells under treatment with or without TJ0113 (5 μM, 48 h) ( n = 3). Results represent the mean ± SD of at least 3 independent experiments. Significance levels are shown within the graphs.
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The mitophagy-driven metabolic–epigenetic axis suppresses SASP factors in senescent NP cells. (A) Western blot analysis of SASP factors and thrombospondin-1 (THBS1) in young and aged NP cells. (B) Schematic illustration of the experimental design. (C) Western blot analysis of SASP factors and THBS1 in aged NP cells treated with vehicle, TJ0113 (5 μM), FX-11(20 μM), or both. (D) Western blot analysis of SASP factors and THBS1 in aged NP cells treated with vehicle, TJ0113 (5 μM), L-Lac (10 μM), or both. (E) qRT-PCR analysis of SASP factors in aged NP cells with treatment as (B) and (C) ( n = 3). (F) Western blot analysis of SASP factors and THBS1 in control and THBS1 knockdown aged NP cells treated with or without TJ0113 (5 μM, 48 h). (G) qRT-PCR analysis of SASP factors in control and THBS1 knockdown aged NP cells treated with TJ0113 or not (5 μM, 48 h) ( n = 3). (H) Western blot analysis of SASP factors and THBS1 in control and THBS1-treated (0.1/1 μg/ml, 48 h) aged NP cells under treatment with or without TJ0113 (5 μM, 48 h). (I) qRT-PCR analysis of SASP factors in control and THBS1-treated (0.1/1 μg/ml, 48 h) aged NP cells under treatment with or without TJ0113 (5 μM, 48 h) ( n = 3). Results represent the mean ± SD of at least 3 independent experiments. Significance levels are shown within the graphs.

Journal: Research

Article Title: Mitophagy Reprograms Lactate Metabolism to Suppress THBS1 via H3K18la Reduction, Alleviating Intervertebral Disc Degeneration

doi: 10.34133/research.0957

Figure Lengend Snippet: The mitophagy-driven metabolic–epigenetic axis suppresses SASP factors in senescent NP cells. (A) Western blot analysis of SASP factors and thrombospondin-1 (THBS1) in young and aged NP cells. (B) Schematic illustration of the experimental design. (C) Western blot analysis of SASP factors and THBS1 in aged NP cells treated with vehicle, TJ0113 (5 μM), FX-11(20 μM), or both. (D) Western blot analysis of SASP factors and THBS1 in aged NP cells treated with vehicle, TJ0113 (5 μM), L-Lac (10 μM), or both. (E) qRT-PCR analysis of SASP factors in aged NP cells with treatment as (B) and (C) ( n = 3). (F) Western blot analysis of SASP factors and THBS1 in control and THBS1 knockdown aged NP cells treated with or without TJ0113 (5 μM, 48 h). (G) qRT-PCR analysis of SASP factors in control and THBS1 knockdown aged NP cells treated with TJ0113 or not (5 μM, 48 h) ( n = 3). (H) Western blot analysis of SASP factors and THBS1 in control and THBS1-treated (0.1/1 μg/ml, 48 h) aged NP cells under treatment with or without TJ0113 (5 μM, 48 h). (I) qRT-PCR analysis of SASP factors in control and THBS1-treated (0.1/1 μg/ml, 48 h) aged NP cells under treatment with or without TJ0113 (5 μM, 48 h) ( n = 3). Results represent the mean ± SD of at least 3 independent experiments. Significance levels are shown within the graphs.

Article Snippet: FX-11 (catalog no. HY-16214), L-Lac (catalog no. HY-Y0479), and THBS1 recombinant protein (catalog no. HY- P70725 ) were purchased from MedChemExpress.

Techniques: Western Blot, Quantitative RT-PCR, Control, Knockdown