Journal: Cell Biochemistry and Function

Article Title: Exosomal let‐7c‐5p From AMI Patient Plasma Modulates Neuronal PANoptosis via ZBP1 In Vitro

doi: 10.1002/cbf.70318

Figure Lengend Snippet: AMI plasma‐derived exosomes are taken up by SH‐SY5Y cells and affect ZBP1 expression and PANoptosis‐related protein levels. (A) Uptake of DiI‐labeled exosomes derived from healthy volunteers and AMI patients (red fluorescence) by SH‐SY5Y cells; (B) Real‐time PCR analysis of ZBP1 mRNA expression and western blot analysis of ZBP1 protein levels; (C) Immunofluorescence staining and quantification of ZBP1 relative fluorescence intensity; (D) Western blot analysis of PANoptosis‐related proteins: Caspase‐8, Caspase‐3, NLRP3, GSDMD, GSDME, RIP3, and MLKL; CCK‐8 assay measuring SH‐SY5Y cell viability following exosome treatment. * p < 0.05 compared with Blank (in all), C (in a, c, i, k), 2 h (in a, d, g, h, j, k) and 24 h (in a, b, c, d, e, g, h, i, j, k) groups; # p < 0.05 compared with Blank (in all), C (in a, c, d) and 24 h (in a, c, e, i, k) groups; & p < 0.05 compared with Blank (in c, d) and C (in c, d, h, j, k) groups. n = 3.

Article Snippet: Human neuroblastoma SH‐SY5Y cells were obtained from the Chinese Academy of Sciences Cell Bank (SCSP‐5012; ATCC CRL‐2266) and authenticated by STR profiling, showing 100% match with the Cellosaurus database (CVCL_0019).

Techniques: Clinical Proteomics, Derivative Assay, Exosomes, Expressing, Labeling, Fluorescence, Real-time Polymerase Chain Reaction, Analysis, Western Blot, Immunofluorescence, Staining, CCK-8 Assay

Journal: Cell Biochemistry and Function

Article Title: Exosomal let‐7c‐5p From AMI Patient Plasma Modulates Neuronal PANoptosis via ZBP1 In Vitro

doi: 10.1002/cbf.70318

Figure Lengend Snippet: Predicted binding of let‐7c‐5p to ZBP1 mRNA and the effects of let‐7c‐5p overexpression on ZBP1 expression and PANoptosis‐related protein levels in SH‐SY5Y cells. (A) Bioinformatic prediction showing two potential binding sites of let‐7c‐5p in ZBP1 mRNA, located within the 3' UTR and 5' UTR. The predicted binding entropy values were 25.2 and 24.5 kcal/mol, respectively, suggesting direct regulation by let‐7c‐5p; (B) GFP fluorescence image of SH‐SY5Y cells 48 h after lentiviral transduction of let‐7c‐5p. RT‐qPCR analysis showing efficient overexpression of let‐7c‐5p; (C) RT‐qPCR analysis of ZBP1 mRNA expression and western blot analysis of ZBP1 protein levels; (D) Western blot analysis of PANoptosis‐related proteins, including Caspase‐3, GSDMD, GSDME, and MLKL. * p < 0.05 versus Blank and C groups. n = 3.

Article Snippet: Human neuroblastoma SH‐SY5Y cells were obtained from the Chinese Academy of Sciences Cell Bank (SCSP‐5012; ATCC CRL‐2266) and authenticated by STR profiling, showing 100% match with the Cellosaurus database (CVCL_0019).

Techniques: Binding Assay, Over Expression, Expressing, Fluorescence, Transduction, Quantitative RT-PCR, Analysis, Western Blot

Journal: Cell Biochemistry and Function

Article Title: Exosomal let‐7c‐5p From AMI Patient Plasma Modulates Neuronal PANoptosis via ZBP1 In Vitro

doi: 10.1002/cbf.70318

Figure Lengend Snippet: Effects of AMI plasma‐derived exosomes on ZBP1 expression and PANoptosis in SH‐SY5Y cells under OGD/R conditions. (A) Real‐time PCR analysis of ZBP1 mRNA expression and western blot analysis of ZBP1 protein levels; (B) Immunofluorescence staining and quantification of ZBP1 relative fluorescence intensity; (C) Western blot analysis of PANoptosis‐related proteins: Caspase‐8, Caspase‐3, NLRP3, GSDMD, GSDME, RIP3, and MLKL; CCK‐8 assay measuring SH‐SY5Y cell viability. * p < 0.05 compared with Blank (in a, c, h, k), C (in a, c, k), OGD‐Blank (in a, b, c, d, e, f), OGD‐2 h (in a, b, c, d, e, g, h, k) and OGD‐24 h (in a, e, j) groups; # p < 0.05compared with Blank (in a, c, d, e, g, h, i, j, k), C (in a, c, e, g, h, k), OGD‐Blank (in c), OGD‐C (in a, c, d, e, g, h, k) and OGD‐24 h (in all) groups; & p < 0.05 compared with Blank (in c, j, k), C (in c, e, f, j), OGD‐Blank (in a, b, c, d, e, f, g, i, j, k) and OGD‐C (in b, e, f, i, j) groups. n = 3.

Article Snippet: Human neuroblastoma SH‐SY5Y cells were obtained from the Chinese Academy of Sciences Cell Bank (SCSP‐5012; ATCC CRL‐2266) and authenticated by STR profiling, showing 100% match with the Cellosaurus database (CVCL_0019).

Techniques: Clinical Proteomics, Derivative Assay, Exosomes, Expressing, Real-time Polymerase Chain Reaction, Analysis, Western Blot, Immunofluorescence, Staining, Fluorescence, CCK-8 Assay