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Impact of SPP1-positive macrophages on the function of <t>msi-mNPCs.</t> Note: ( A ) Correlation analysis of SPP1 and Macrophage abundance in RNA-seq data; ( B ) Western blot detection of SPP1 expression in Control and msi-IDD mouse IVD tissues; ( C ) Influence of co-culture with oe-NC and oe-SPP1 on the proliferation capacity of msi-mNPCs; ( D ) Influence of co-culture with oe-NC and oe-SPP1 on the apoptosis level of msi-mNPCs, with representative images on the left and quantification on the right; ( E – G ) Effect of oe-NC and oe-SPP1 on the secretion of extracellular matrix and inflammatory factors in msi-mNPCs detected by ELISA; ( H - I ) Influence of co-culture with sh-NC and sh-SPP1 on the proliferation and apoptosis of msi-mNPCs; ( J - L ) Effect of sh-NC and sh-SPP1 on the secretion of extracellular matrix and inflammatory factors in msi-mNPCs detected by ELISA; ( M – N ) Western blot detection of the impact of oe-SPP1 or sh-SPP1 on the expression of msi-mNPC-related factors; All cell experiments were repeated three times; ns P > 0.05, ** P < 0.01, *** P < 0.001
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Impact of SPP1-positive macrophages on the function of <t>msi-mNPCs.</t> Note: ( A ) Correlation analysis of SPP1 and Macrophage abundance in RNA-seq data; ( B ) Western blot detection of SPP1 expression in Control and msi-IDD mouse IVD tissues; ( C ) Influence of co-culture with oe-NC and oe-SPP1 on the proliferation capacity of msi-mNPCs; ( D ) Influence of co-culture with oe-NC and oe-SPP1 on the apoptosis level of msi-mNPCs, with representative images on the left and quantification on the right; ( E – G ) Effect of oe-NC and oe-SPP1 on the secretion of extracellular matrix and inflammatory factors in msi-mNPCs detected by ELISA; ( H - I ) Influence of co-culture with sh-NC and sh-SPP1 on the proliferation and apoptosis of msi-mNPCs; ( J - L ) Effect of sh-NC and sh-SPP1 on the secretion of extracellular matrix and inflammatory factors in msi-mNPCs detected by ELISA; ( M – N ) Western blot detection of the impact of oe-SPP1 or sh-SPP1 on the expression of msi-mNPC-related factors; All cell experiments were repeated three times; ns P > 0.05, ** P < 0.01, *** P < 0.001
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Impact of SPP1-positive macrophages on the function of msi-mNPCs. Note: ( A ) Correlation analysis of SPP1 and Macrophage abundance in RNA-seq data; ( B ) Western blot detection of SPP1 expression in Control and msi-IDD mouse IVD tissues; ( C ) Influence of co-culture with oe-NC and oe-SPP1 on the proliferation capacity of msi-mNPCs; ( D ) Influence of co-culture with oe-NC and oe-SPP1 on the apoptosis level of msi-mNPCs, with representative images on the left and quantification on the right; ( E – G ) Effect of oe-NC and oe-SPP1 on the secretion of extracellular matrix and inflammatory factors in msi-mNPCs detected by ELISA; ( H - I ) Influence of co-culture with sh-NC and sh-SPP1 on the proliferation and apoptosis of msi-mNPCs; ( J - L ) Effect of sh-NC and sh-SPP1 on the secretion of extracellular matrix and inflammatory factors in msi-mNPCs detected by ELISA; ( M – N ) Western blot detection of the impact of oe-SPP1 or sh-SPP1 on the expression of msi-mNPC-related factors; All cell experiments were repeated three times; ns P > 0.05, ** P < 0.01, *** P < 0.001

Journal: Cell Biology and Toxicology

Article Title: Deciphering SPP1-related macrophage signaling in the pathogenesis of intervertebral disc degeneration

doi: 10.1007/s10565-024-09948-4

Figure Lengend Snippet: Impact of SPP1-positive macrophages on the function of msi-mNPCs. Note: ( A ) Correlation analysis of SPP1 and Macrophage abundance in RNA-seq data; ( B ) Western blot detection of SPP1 expression in Control and msi-IDD mouse IVD tissues; ( C ) Influence of co-culture with oe-NC and oe-SPP1 on the proliferation capacity of msi-mNPCs; ( D ) Influence of co-culture with oe-NC and oe-SPP1 on the apoptosis level of msi-mNPCs, with representative images on the left and quantification on the right; ( E – G ) Effect of oe-NC and oe-SPP1 on the secretion of extracellular matrix and inflammatory factors in msi-mNPCs detected by ELISA; ( H - I ) Influence of co-culture with sh-NC and sh-SPP1 on the proliferation and apoptosis of msi-mNPCs; ( J - L ) Effect of sh-NC and sh-SPP1 on the secretion of extracellular matrix and inflammatory factors in msi-mNPCs detected by ELISA; ( M – N ) Western blot detection of the impact of oe-SPP1 or sh-SPP1 on the expression of msi-mNPC-related factors; All cell experiments were repeated three times; ns P > 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The mouse nucleus pulposus cells (mNPCs) (CP-M146, China, provided by Procell) were cultured in Thermo Fisher's DMEM/F-12 (11320033, USA).

Techniques: RNA Sequencing, Western Blot, Expressing, Control, Co-Culture Assay, Enzyme-linked Immunosorbent Assay

Impact of the SPP1/PERK/ATF4/IL-10 signaling axis on msi-mNPCs functionality. Note: ( A ) Effects of GSK2656157 treatment on the expression and activation of sh-SPP1-related factors. ( B ) Effect of GSK2656157-treated sh-SPP1 on msi-mNPCs proliferation detected by CCK8 assay. ( C ) Effect of GSK2656157-treated sh-SPP1 on the secretion of Collagen Iα1 and Collagen II in msi-mNPCs detected by ELISA. ( D ) Effect of ATF4 knockdown on sh-SPP1-induced msi-mNPCs proliferation. ( E – F ) Effects of ATF4 knockdown sh-SPP1 on msi-mNPCs proliferation and the secretions of Collagen Iα1 and Collagen II. ( G ) Effects of anti-IL-10 treatment on the expression and activation of sh-SPP1-related factors. ( H - I ) Effects of anti-IL-10 treatment on msi-mNPCs proliferation and the secretions of Collagen Iα1 and Collagen II. All cell experiments were repeated three times. ns P > 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001

Journal: Cell Biology and Toxicology

Article Title: Deciphering SPP1-related macrophage signaling in the pathogenesis of intervertebral disc degeneration

doi: 10.1007/s10565-024-09948-4

Figure Lengend Snippet: Impact of the SPP1/PERK/ATF4/IL-10 signaling axis on msi-mNPCs functionality. Note: ( A ) Effects of GSK2656157 treatment on the expression and activation of sh-SPP1-related factors. ( B ) Effect of GSK2656157-treated sh-SPP1 on msi-mNPCs proliferation detected by CCK8 assay. ( C ) Effect of GSK2656157-treated sh-SPP1 on the secretion of Collagen Iα1 and Collagen II in msi-mNPCs detected by ELISA. ( D ) Effect of ATF4 knockdown on sh-SPP1-induced msi-mNPCs proliferation. ( E – F ) Effects of ATF4 knockdown sh-SPP1 on msi-mNPCs proliferation and the secretions of Collagen Iα1 and Collagen II. ( G ) Effects of anti-IL-10 treatment on the expression and activation of sh-SPP1-related factors. ( H - I ) Effects of anti-IL-10 treatment on msi-mNPCs proliferation and the secretions of Collagen Iα1 and Collagen II. All cell experiments were repeated three times. ns P > 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The mouse nucleus pulposus cells (mNPCs) (CP-M146, China, provided by Procell) were cultured in Thermo Fisher's DMEM/F-12 (11320033, USA).

Techniques: Expressing, Activation Assay, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Knockdown