antibodies recombinant thbs1  (R&D Systems)

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: Down-regulation of lactate production and histone lactylation is driven by mitophagy activation. (A) Lactate levels of the young and aged NP cells treated with or without TJ0113 (5 μM, 48 h; n = 5) after knocking down MCL1. (B) Western blot analysis of PAN-KLA in young and aged NP cells treated with or without TJ0113 (5 μM, 48 h) after knocking down MCL1. (C and D) Western blot analysis of lysine lactylation markers (H3K9la, H3K14la, H3K18la, H4K12la) treated with TJ0113 or not (5 μM, 48 h). Histone H3/H4 served as loading controls. (E) Heatmap depicting H3K18la genomic occupancy profiles [±3 kb flanking transcription start sites (TSSs)] treated with TJ0113 or not (5 μM, 48 h). Genes are ranked vertically by descending H3K18la signal intensity. (F) Comparative pie charts showing genome-wide differential H3K18la distribution patterns at annotated genomic regions in aged NP cells treated with TJ0113 (5 μM, 48 h) compared to untreated controls. (G) KEGG enrichment analysis of H3K18la-marked genes. (H) Venn diagram depicting genes down-regulated in RNA-seq analysis of aged NP cells treated with TJ0113 (5 μM, 48 h) compared to untreated controls and the down-regulated target genes bound by H3K18la under identical treatment conditions. (I) Normalized read densities for H3K18la at the THBS1 genes. (J) ChIP-qPCR assay in aged NP cells. 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: Activation Assay, Western Blot, Genome Wide, RNA Sequencing, ChIP-qPCR

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

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: Targeting the lactate–H3K18la–THBS1 axis attenuates IVDD. (A) Schematic illustration of the experimental design. (B) Representative images of H&E and SO&FG staining of rat coccygeal IVDs at 12 weeks after treatment. Scale bar, 1 mm. (C) histological score ( n = 6) of rat coccygeal IVDs. (D) Representative immunohistochemistry images of H3K18la in rat coccygeal IVDs at 12 weeks after treatment. Scale bar, 200 μm. (E) Representative IF images of THBS1 in rat coccygeal IVDs at 12 weeks after treatment. Scale bar, 500 μm. (F and G) Quantification of SASP factors in rat coccygeal IVDs. 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: Staining, Immunohistochemistry