Journal: Cellular & Molecular Biology Letters

Article Title: PCSK9 inhibition ameliorates microplastic-induced endothelial redox imbalance via SIRT6 modulation

doi: 10.1186/s11658-025-00838-z

Figure Lengend Snippet: Effects of MPs on teloHAEC. Cell viability evaluated in teloHAEC exposed to different concentrations of A PE and B PVC (0–70 µg/mL) for 24, 48, and 72 h. C , D Cell viability assessed after 48 h of treatment with PE or PVC (30 µg/mL) and increasing concentration of PVC or PE (0–70 µg/mL), respectively. Evaluation of E MCP-1, F VCAM1, and G ICAM1 content in teloHAEC treated with PE (70 µg/mL), PVC (70 µg/mL), or PE + PVC (30 µg/mL + 30 µg/mL) for 48 h. H , I Representative cell cycle detection by FACS analysis in EC exposed to MPs. Data expressed as mean ± standard deviation (SD) of n = 4 independent experiments. * p < 0.05 versus 0 µg/mL or Ctr; ** p < 0.01 versus 0 µg/mL or Ctr; • p < 0.001 versus 0 µg/mL or Ctr; † p < 0.05 versus PE; ‡ p < 0.05 versus PVC; ns, not significant versus PE and PVC

Article Snippet: The levels of inflammatory mediators MCP-1 (RAF081R, BioVendor, Brno, Czech Republic), VCAM1 (EH0326, FineTest, Hubei, China), and ICAM1 (EH0161, FineTest, Hubei, China) were determined in cell culture supernatant, as previously reported [ ].

Techniques: Concentration Assay, Standard Deviation

Journal: Cellular & Molecular Biology Letters

Article Title: PCSK9 inhibition ameliorates microplastic-induced endothelial redox imbalance via SIRT6 modulation

doi: 10.1186/s11658-025-00838-z

Figure Lengend Snippet: iPCSK9 opposed the MP-related inflammation. A TeloHAEC viability evaluated after treatment with PE and PVC alone or combined PE + PVC, or pretreated with iPCSK9 (100 ng/mL) for 8 h and then exposed to PE, PVC, and PE + PVC. B Immunoblotting analysis of SIRT6 protein levels and ELISA assays of C MCP-1, D VCAM1, and E ICAM1. F Representative annexin V-FITC and PI-staining detected by FACS analysis and G cell cycle investigation. Data expressed as mean ± SD of n = 4 experiments. M, molecular weight markers; lane 1, Ctr; lane 2, PE; lane 3, PVC; lane 4, PE + PVC; lane 5, iPCSK9; lane 6, PE + iPCSK9; lane 7, PVC + iPCSK9; lane 8, PE + PVC + iPCSK9. * p < 0.05 versus Ctr; ** p < 0.01 versus Ctr; • p < 0.001 versus Ctr; † p < 0.05 versus PE; ‡ p < 0.05 versus PVC; + p < 0.05 versus PE + PVC; ¶ p < 0.01 versus PE; § p < 0.01 versus PVC; ▲ p < 0.01 versus PE + PVC; ns, not significant versus PE and PVC

Article Snippet: The levels of inflammatory mediators MCP-1 (RAF081R, BioVendor, Brno, Czech Republic), VCAM1 (EH0326, FineTest, Hubei, China), and ICAM1 (EH0161, FineTest, Hubei, China) were determined in cell culture supernatant, as previously reported [ ].

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Molecular Weight

Journal: Cellular & Molecular Biology Letters

Article Title: PCSK9 inhibition ameliorates microplastic-induced endothelial redox imbalance via SIRT6 modulation

doi: 10.1186/s11658-025-00838-z

Figure Lengend Snippet: SIRT6 silencing opposed the effects of iPCSK9 on inflammation and apoptosis. A Immunoblotting analysis of SIRT6 protein levels in teloHAEC treated with the empty transfection reagent (Vehicle) or transfected with negative control siRNA (NT) or with SIRT6 siRNA (siSIRT6). M, molecular weight markers; lane 1, Ctr; lane 2, Vehicle; lane 3, NT; lane 4, siSIRT6. B Immunoblotting analysis of PCSK9 protein levels and ELISA assays of C MCP-1, D VCAM1 and E ICAM1. F , G Representative dot plots and FACS analysis of annexin V-FITC and PI-staining of EC transfected with NT or siSIRT6 and exposed to PE + PVC alone or pretreated with iPCSK9 (100 µg/mL) before MP stimulation. M, molecular weight markers; lane 1, Ctr; lane 2, NT; lane 3, NT + PE + PVC; lane 4, NT + iPCSK9; lane 5, NT + PE + PVC + iPCSK9; lane 6, siSIRT6; lane 7, siSIRT6 + PE + PVC; lane 8, siSIRT6 + iPCSK9; lane 9, siSIRT6 + PE + PVC + iPCSK9. Q1: necrotic cells; Q2: late apoptotic cells; Q3: early apoptotic cells; Q4: viable cells. Data are expressed as mean ± SD of n = 3 experiments. ° p < 0.01 versus NT; # p < 0.001 versus NT; + p < 0.05 versus PE + PVC; ▲ p < 0.01 versus PE + PVC

Article Snippet: The levels of inflammatory mediators MCP-1 (RAF081R, BioVendor, Brno, Czech Republic), VCAM1 (EH0326, FineTest, Hubei, China), and ICAM1 (EH0161, FineTest, Hubei, China) were determined in cell culture supernatant, as previously reported [ ].

Techniques: Western Blot, Transfection, Negative Control, Molecular Weight, Enzyme-linked Immunosorbent Assay, Staining

Journal: Journal for Immunotherapy of Cancer

Article Title: Disruption of tumor-intrinsic PGAM5 increases anti-PD-1 efficacy through the CCL2 signaling pathway

doi: 10.1136/jitc-2024-009993

Figure Lengend Snippet: PGAM5 reduced CCL2 secretion in HCC cells by downregulating DRP1. ( A ) Schematic diagram of the in vitro co-culture system for Huh7 shctrl or shPGAM5 cells and phorbol-12-myristate-13-acetate-induced THP-1 cells. ( B ) Quantitative PCR analysis of macrophage polarization maker genes of THP-1 cells co-cultured with indicated HCC cells. ( C ) cytokine antibody array incubated with culture medium of Huh7 shctrl or shPGAM5 cells. ( D ) Quantitative PCR analysis of macrophage M1/M2 polarization maker genes of THP-1 in the co-culture system, culture medium of Huh7 shPGAM5 cells were supplemented with recombinant CCL2 and HCCLM3 PGAM5 OE cells were supplemented with neutralizing-CCL2 antibody. ( E ) TOP20 GO enrichment of differentially expressed proteins in Huh7 shPGAM5 cells compared with shctrl cells. ( F ) Volcano plot of significantly differentially expressed proteins in Huh7 shPGAM5 cells compared with shctrl cells. ( G ) Western blot analysis of protein expression of PGAM5 and mitochondria dynamic proteins DRP1, MFN1, MFN2, OPA1 in HCC cells. ( H ) Western blot analysis of DRP1 overexpression efficiency in Huh7 shPGAM5 cells by lenti-virus infection. ( I ) Concentration of CCL2 in the supernatant of indicated HCC cells was measured by ELISA. ( J ) Western blot analysis of DRP1 silencing efficiency in HCCLM3 PGAM5-OE cells by transient transfection of SiRNA. ( K ) Concentration of CCL2 in the supernatant of indicated HCC cells was measured by ELISA kit. *p<0.05, **p<0.01, and ***p<0.001, ****p<0.0001, ns: not significant. HCC, hepatocellular carcinoma; IHC, immunohistochemistry; UMAP, uniform manifold approximation and projection.

Article Snippet: Concentrations of CCL2 in the supernatant were examined by ELISA kit (CSB-E04655h, cusabio) following manufacturer’s instructions.

Techniques: In Vitro, Co-Culture Assay, Real-time Polymerase Chain Reaction, Cell Culture, Ab Array, Incubation, Recombinant, Western Blot, Expressing, Over Expression, Virus, Infection, Concentration Assay, Enzyme-linked Immunosorbent Assay, Transfection, Immunohistochemistry

Journal: Regenerative Biomaterials

Article Title: AI-guided design and optimization of a novel KIM-1-targeted peptide for bFGF delivery in acute kidney injury repair

doi: 10.1093/rb/rbag050

Figure Lengend Snippet: Construction and bioactivity evaluation of WEV-bFGF. ( A ) Spatial structure prediction of WEV-bFGF recombinant protein and natural bFGF; ( B ) SDS-PAGE of bFGF, KIT-bFGF and WEV-bFGF; ( C ) Western blot of native bFGF, KIT-bFGF and WEV-bFGF; ( D ) The biological activities of native bFGF, KIT-bFGF and WEV-bFGF by CCK-8 assay; ( E ) The fluorescence distribution of Delight 800 labeled recombinant proteins in hypoxic HK-2 cells; ( F ) Statistical analysis of fluorescence intensity of Dylight 800 in hypoxic HK-2 cells; Scale bar = 25 μm; ( G ) The protective effects of recombinant proteins on HK-2 cells after hypoxic injury by the CCK8 assay. All quantitative data are presented as mean ± SD, * P < 0.05, ** P < 0.01.

Article Snippet: Subsequently, in accordance with the instructions of the Human bFGF ELISA Kit (EK0441, Boster, China), the protein extracted from kidney tissue and the content of bFGF in serum were determined.

Techniques: Recombinant, SDS Page, Western Blot, CCK-8 Assay, Fluorescence, Labeling

Journal: Regenerative Biomaterials

Article Title: AI-guided design and optimization of a novel KIM-1-targeted peptide for bFGF delivery in acute kidney injury repair

doi: 10.1093/rb/rbag050

Figure Lengend Snippet: The evaluation of targeting capacity of WEV-bFGF in ischemic kidney in vivo . ( A ) At 6 h after administration, the in vivo fluorescence distribution of Dylight800-labeled recombinant proteins in major organs of AKI-injured rats by the small animal imaging system; ( B ) Western blot showing bFGF protein expression in ischemic renal tissue 6 h after administration; ( C ) Quantitative analysis of relative bFGF protein expression at 6 h; ( D ) The content of bFGF in ischemic kidneys by ELISA assay at 6 h after administration; ( E ) The content of bFGF in serum at 6 h after administration; ( F ) Immunofluorescence colocalization of bFGF and KIM-1 in the frozen sections at 6 h after administration, Scale bar = 50 μm; ( G ) Colocalization analysis of PBS, bFGF, KIT-bFGF and WEV-bFGF groups at 6 h after administration; ( H ) At 24 h after administration, the in vivo fluorescence distribution of Dylight800-labeled recombinant proteins in major organs of AKI-injured rats by the small animal imaging system; ( I ) Western blot showing bFGF protein expression in ischemic renal tissue 24 h after administration; ( J ) Quantitative analysis of relative bFGF protein expression at 24 h; ( K ) The content of bFGF in ischemic kidneys by ELISA assay at 24 h after administration; ( L ) The content of bFGF in serum at 24 h after administration; ( M ) Immunofluorescence staining showing colocalization of bFGF and KIM-1 at 24 h after administration, scale bar = 50 μm; ( N ) Colocalization analysis of PBS, bFGF, KIT-bFGF and WEV-bFGF groups at 24 h after administration. All quantitative data are presented as mean ± SD, * P < 0.05, ** P < 0.01, N = 8.

Article Snippet: Subsequently, in accordance with the instructions of the Human bFGF ELISA Kit (EK0441, Boster, China), the protein extracted from kidney tissue and the content of bFGF in serum were determined.

Techniques: In Vivo, Fluorescence, Labeling, Recombinant, Imaging, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining

Journal: Regenerative Biomaterials

Article Title: AI-guided design and optimization of a novel KIM-1-targeted peptide for bFGF delivery in acute kidney injury repair

doi: 10.1093/rb/rbag050

Figure Lengend Snippet: Functional tests and morphological analysis of the kidneys. ( A ) Experimental design of WEV-bFGF for the treatment of I/R; ( B ) Statistical analysis of serum Scr content for renal function evaluation at 24 h after renal I/R injury in rats; ( C ) Statistical analysis of serum Scr content for renal function evaluation at 24 h after renal I/R injury in rats; ( D ) H&E staining for kidney histopathological evaluation after I/R injury. Arrows indicated the sites of renal tubular injury. Scale bar = 20 μm; ( E ) Statistical analysis of renal tubular injury score; ( F ) MASSON staining for renal tissue fibrosis evaluation after I/R injury. Scale bar = 50 μm; ( G ) Statistical analysis of renal tissue fibrosis. All quantitative data are presented as mean ± SD, * P < 0.05, ** P < 0.01, N = 8.

Article Snippet: Subsequently, in accordance with the instructions of the Human bFGF ELISA Kit (EK0441, Boster, China), the protein extracted from kidney tissue and the content of bFGF in serum were determined.

Techniques: Functional Assay, Staining

Journal: Regenerative Biomaterials

Article Title: AI-guided design and optimization of a novel KIM-1-targeted peptide for bFGF delivery in acute kidney injury repair

doi: 10.1093/rb/rbag050

Figure Lengend Snippet: Transcriptome analysis of potential mechanism in WEV-bFGF mediating ischemic renal repair. ( A ) Volcano plots of gene differences between the WEV group and the PBS group; ( B ) KEGG analysis between the WEV group and the PBS group; ( C ) Volcano plots of gene differences between the WEV-BFGF group and the bFGF group; ( D ) KEGG analysis between the WEV-bFGF group and the bFGF group; ( E ) Heat map of differentially expressed genes related to repair and regeneration of AKI; ( F ) Quantitative PCR validation of key gene expression. All quantitative data are presented as mean ± SD, * P < 0.05, ** P < 0.01, N = 6.

Article Snippet: Subsequently, in accordance with the instructions of the Human bFGF ELISA Kit (EK0441, Boster, China), the protein extracted from kidney tissue and the content of bFGF in serum were determined.

Techniques: Real-time Polymerase Chain Reaction, Biomarker Discovery, Gene Expression