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

Journal: Nature Communications

Article Title: IL-17A is increased in diabetic wounds and impairs keratinocyte function via histone demethylase JMJD3

doi: 10.1038/s41467-025-67456-3

Figure Lengend Snippet: a ChIP analysis of H3K27me3 or IgG at indicated promoters in keratinocytes with (white) or without (blue) IL-17A stimulation. Itga3: n = 5 (unstimulated), n = 4 (IL-17A-stimulated) technical replicates, p = 0.0034, Timp1 : n = 3 technical replicates, p = 0.0046, Ccl20 : n = 3 technical replicates, p = 0.0059, Cxcl1 : n = 3 technical replicates, p = 0.0020, Cxcl3 : n = 3 technical replicates, p = 0.0258, Cxcl5 : n = 3 technical replicates, p = 0.0174. n = 3 independent experiments. b qPCR analysis of keratinocytes treated with DMSO only (white), with IL-17A alone (blue), or with IL-17A and GSK-J4 (1 µM) (red). Itga3 : n = 4 biological replicates, p < 0.0001 (DMSO vs. IL-17A), p = 0.0017 (IL-17A vs. IL-17A and inhibitor), Timp1: n = 6 biological replicates, p < 0.0312 (DMSO vs. IL-17A), p = 0.0029 (IL-17A vs. IL-17A and inhibitor), Ccl20: n = 3 biological replicates, p = 0.0008 (DMSO vs. IL-17A), p = 0.0428 (IL-17A vs. IL-17A and inhibitor), Cxcl1 : n = 3 biological replicates, p = 0.0003 (DMSO vs. IL-17A), p = 0.0294 (IL-17A vs. IL-17A and inhibitor), Cxcl3 : n = 3 biological replicates, p < 0.0001 (DMSO vs. IL-17A), p = 0.0003 (IL-17A vs. IL-17A and inhibitor), Cxcl5 : n = 3 biological replicates, p < 0.0001 (DMSO vs. IL-17A), p = 0.0007 (IL-17A vs. IL-17A and inhibitor), n = 3 independent experiments. c Western blot of ITGA-3 expression in keratinocytes treated with DMSO only (white), with IL-17A alone (blue), or with IL-17A and GSK-J4 (red). Representative densitometry plot is shown. n = 3 independent experiments. d Protein quantification of lysates from keratinocytes treated with DMSO only (white), with IL-17A alone (blue), or with IL-17A and GSK-J4 (red). TIMP-1: n = 6 biological replicates, p = 0.0012 (DMSO vs. IL-17A), p = 0.0068 (IL-17A vs. IL-17A and inhibitor), CCL-20: n = 3 biological replicates, p < 0.0001 (DMSO vs. IL-17A), p = 0.0199 (IL-17A vs. IL-17A and inhibitor), CXCL-1: n = 3 biological replicates, p < 0.0001 (DMSO vs. IL-17A), p = 0.0158 (IL-17A vs. IL-17A and inhibitor), CXCL-3: n = 3 biological replicates, p < 0.0001 (DMSO vs. IL-17A), p = 0.0167 (IL-17A vs. IL-17A and inhibitor), CXCL-5: n = 3 biological replicates, p < 0.0001 (DMSO vs. IL-17A), p = 0.0005 (IL-17A vs. IL-17A and inhibitor), n = 3 independent experiments. e qPCR analysis of keratinocytes treated with a non-targeting control (siNTC) (white) or si Jmjd3 (gray). Jmjd3 : n = 6 (siNTC), n = 4 biological replicates (si Jmjd3 ), p = 0.0192, Itga3: n = 6 (siNTC), n = 4 biological replicates (si Jmjd3 ), p = 0.0019, Timp1: n = 3 (siNTC), n = 4 biological replicates (si Jmjd3 ), p = 0.0205, Ccl20: n = 6 (siNTC), n = 4 biological replicates (si Jmjd3 ), p = 0.0015, Cxcl1: n = 6 (siNTC), n = 4 biological replicates (si Jmjd3 ), p = 0.0019, Cxcl3: n = 6 (siNTC), n = 4 biological replicates (si Jmjd3 ), p = 0.0011, Cxcl5: n = 6 (siNTC), n = 4 biological replicates (si Jmjd3 ), p = 0.0011, n = 3 independent experiments. f qPCR analysis of Jmjd3 fl/fl K14 cre+ (red) and Jmjd3 fl/fl K14 cre- (yellow) keratinocytes. n = 3 biological replicates, Itga3 : p = 0.0184, Timp1 : p = 0.0044, Ccl20 : p = 0.0160, Cxcl1 : p = 0.0371, Cxcl3 : p = 0.0042. n = 3 independent experiments. g , h Scratch assays of primary murine ( n = 3 biological replicates, p < 0.0001 (48 h)) and N/TERT ( n = 3 biological replicates, p = 0.0340 (12 h)) keratinocytes treated with IL-17A alone (blue) or IL-17A and GSK-J4 (red). n = 3 independent experiments. i , j Scratch assays of primary murine ( n = 3 biological replicates, p = 0.0016 (48 h)) and N/TERT ( n = 6 biological replicates (IL-17A alone), n = 4 biological replicates (IL-17A and GSK-J1), p = 0.0223 (8 h), p < 0.0001 (12 h)) keratinocytes treated with IL-17A alone (blue) or IL-17A and GSK-J1 (red). n = 3 independent experiments. k Scratch assay of Jmjd3 fl/fl K14 cre+ (red) and Jmjd3 fl/fl K14 cre- (blue) keratinocytes. n = 3 biological replicates, p = 0.0198 (48 h). n = 3 independent experiments. Data were analyzed for variances, and 2-tailed Student’s t tests for ( a ), ( e ), ( f ) and 1-way ANOVA tests for ( b ), ( d ), ( g – k ) were performed. Data are presented as the mean ± SEM.

Article Snippet: After stimulation, cell free supernatant was collected and analyzed by the University of Michigan Immune Monitoring Shared Resource Core for CCL-20, CXCL-1, CXCL-5 or specific enzyme immunoassay kits for CXCL-3 (Boster Bio) and TIMP-1 (R&D Systems) according to the manufacturer’s instructions.

Techniques: Western Blot, Expressing, Control, Wound Healing Assay

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Antagonist Antibodies to PD-1 and B7-H1 (PD-L1) in the Treatment of Advanced Human Cancer

doi: 10.1158/1078-0432.CCR-12-2063

Figure Lengend Snippet: PD-1, B7-H1, and B7-DC expression and prognostic significance in cancer patients

Article Snippet: , ( 58 ) , 150 , Paraffin IHC; anti-B7-H1 mAb (5H1) or anti- B7-H1 polyclonal Ab (4059, ProSci) , Membranous PD-L1 expression by melanocytes within the tumors had 3 patterns: no PD-L1; regional expression of PD-L1 on melanocytes colocalized with TILs (most common); and PD-L1 expression absence of TILs , Almost all PD-L1+ tumors were associated with TILs, while only 28% of PD-L1- tumors were associated with TILs; a sample of PD-L1+ tumors with TIL were shown to contain IFNγ, whereas no PD-L1- tumors examined contained IFNγ , PD-L1 was expressed on a proportion (57/150) of various MEL lesions, most commonly in close juxtaposition to TILs; when an inflammatory response to the tumor was detected, it was likely that both the tumor and infiltrating cells were PD-L1+; PD-L1 expression was associated with the superficial spreading and nodular MEL subtypes ( P =0.033) and not with MEL stage , Patients with PD-L1+ metastatic MEL (mMEL) had longer survival than those with PD-L1- metastatic MEL ( P = 0.032); patients with metastatic MEL with TILs had significantly improved survival compared with those without TILs ( P = 0.017).

Techniques: Expressing, Clone Assay, Membrane, Clinical Proteomics, Staining, Comparison, Functional Assay, Western Blot