Journal: Scientific Reports

Article Title: YBX1/PPIB axis promotes post-maturation arteriovenous fistula stenosis via enhancing endothelial to mesenchymal transition

doi: 10.1038/s41598-026-49568-y

Figure Lengend Snippet: Identification of AVFS-Associated Cell Subpopulations and Candidate Genes. (A) UMAP plot showing the grouping of 10 cell types. (B) Expression of key marker genes across cell types. (C) Differential proportions of each cell type between AVFS and non-stenotic control. (D) Volcano plot of DEGs in endothelial cells. (E-G) GO, KEGG and Reactome enrichment analysis illustrating predominant biological functions and pathways. (H) Flow cytometry sorting and analysis of endothelial cells from vascular tissues, n = 3. (I) qRT-PCR analysis of PPIB, CD31, VE-cadherin, and α-SMA expression levels in endothelial cells from AVFS and non-stenosis tissues, n = 3. (J) WB analysis of PPIB, CD31, VE-cadherin, and α-SMA expression levels in endothelial cells from AVFS and non-stenosis tissues, n = 3. Data are presented as mean ± SD, differences between groups were analyzed by paired t-test. * P < 0.05; **<0.01; ***<0.001; ****<0.0001. SMC: Smooth muscle cell, EC: Endothelial cell, GO: Gene Ontology, KEGG: Kyoto Encyclopedia of Genes and Genomes.

Article Snippet: The antibodies used included: anti-PPIB antibody (CUSABIO, China; #CSB-PA018472ESR1HU), anti-YBX1 antibody (Abclonal, China; #A3534), anti-VE-cadherin antibody (Abclonal, China; #A12416), anti-CD31 antibody (Abclonal, China; #A19014), anti-α-SMA antibody (Affinity, China, #AF1032), and anti-GAPDH antibody (Abclonal, China; #A19056).

Techniques: Expressing, Marker, Control, Flow Cytometry, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: YBX1/PPIB axis promotes post-maturation arteriovenous fistula stenosis via enhancing endothelial to mesenchymal transition

doi: 10.1038/s41598-026-49568-y

Figure Lengend Snippet: PPIB Regulates Endothelial Marker in HUVECs. (A) qRT-PCR of PPIB, VE-cadherin, and CD31 expression following PPIB knockdown in HUVECs ( n = 3). (B) qRT-PCR analysis of PPIB, VE-cadherin, and CD31 expression following PPIB overexpression in HUVECs ( n = 3). (C) Western blot of PPIB, VE-cadherin and CD31 after PPIB knockdown in HUVECs ( n = 3). (D) Western blot of PPIB, VE-cadherin, and CD31 expression after PPIB overexpression in HUVECs ( n = 3). The protein bands of WB were derived from different gels, and the uncropped original WB images are provided in the Supplementary Material 1. Data are presented as mean ± SD, differences between groups were analyzed by one-way ANOVA, statistical significance was determined using one-way ANOVA. Compared to the control group, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; compared to the TGF-β1 group, # P < 0.05, ## P < 0.01, ### P < 0.001, #### P < 0.0001. sh: short hairpin, gene expression interference by introducing short hairpin RNA. oe: overexpression, increasing gene expression by introducing the target gene sequence. NC: Negative control, for sh, refers to lentivirus construction by transfecting plasmids with a scrambled shRNA sequence; for oe, refers to lentivirus construction by directly transfecting empty plasmids. ns: not significant.

Article Snippet: The antibodies used included: anti-PPIB antibody (CUSABIO, China; #CSB-PA018472ESR1HU), anti-YBX1 antibody (Abclonal, China; #A3534), anti-VE-cadherin antibody (Abclonal, China; #A12416), anti-CD31 antibody (Abclonal, China; #A19014), anti-α-SMA antibody (Affinity, China, #AF1032), and anti-GAPDH antibody (Abclonal, China; #A19056).

Techniques: Marker, Quantitative RT-PCR, Expressing, Knockdown, Over Expression, Western Blot, Derivative Assay, Control, Gene Expression, shRNA, Sequencing, Negative Control

Journal: Scientific Reports

Article Title: YBX1/PPIB axis promotes post-maturation arteriovenous fistula stenosis via enhancing endothelial to mesenchymal transition

doi: 10.1038/s41598-026-49568-y

Figure Lengend Snippet: Transcription Factor YBX1 is Associated with EndMT in Endothelial Cells. (A) Prediction of YBX1 binding sites in the PPIB promoter and schematic of site-directed mutations used in the dual-luciferase assay. (B) Dual-luciferase assay validating the interaction between YBX1 and the PPIB promoter region ( n = 3). (C) qRT-PCR analysis of YBX1, PPIB, VE-cadherin, and CD31 expression levels ( n = 3). (D–E) Western blot of YBX1, PPIB, VE-cadherin, and CD31 expression levels ( n = 3). (F , H) Immunofluorescence staining assessing α-SMA expression levels ( n = 3). (G , I) Wound healing assays evaluating HUVEC migration ability ( n = 3). The protein bands of WB were derived from different gels, and the uncropped original WB images are provided in the Supplementary Material 1. Data are presented as mean ± SD, differences between groups were analyzed by one-way ANOVA, statistical significance was determined using one-way ANOVA. * P < 0.05, **< 0.01, ***< 0.001, ****< 0.0001. luc: Firefly luciferase. RL: Renilla luciferase. WT: Wild type. Mut: Mutant. oe: overexpression, increasing gene expression by introducing the target gene sequence. NC: Negative control, for oe, refers to lentivirus construction by directly transfecting empty plasmids. ns: not significant.

Article Snippet: The antibodies used included: anti-PPIB antibody (CUSABIO, China; #CSB-PA018472ESR1HU), anti-YBX1 antibody (Abclonal, China; #A3534), anti-VE-cadherin antibody (Abclonal, China; #A12416), anti-CD31 antibody (Abclonal, China; #A19014), anti-α-SMA antibody (Affinity, China, #AF1032), and anti-GAPDH antibody (Abclonal, China; #A19056).

Techniques: Binding Assay, Luciferase, Quantitative RT-PCR, Expressing, Western Blot, Immunofluorescence, Staining, Migration, Derivative Assay, Mutagenesis, Over Expression, Gene Expression, Sequencing, Negative Control

Journal: Scientific Reports

Article Title: YBX1/PPIB axis promotes post-maturation arteriovenous fistula stenosis via enhancing endothelial to mesenchymal transition

doi: 10.1038/s41598-026-49568-y

Figure Lengend Snippet: YBX1 Promotes EndMT through Regulation of PPIB. (A) qRT-PCR assessed the expression levels of YBX1, PPIB, VE-cadherin, and CD31 in each group, n = 3. (B) Western blot assessed the expression levels of YBX1, PPIB, VE-cadherin, and CD31 in each group, n = 3. (C) Immunofluorescence staining of α-SMA expression in HUVECs from each group ( n = 3). (D) Wound healing assay evaluating HUVEC migratory capacity in each group ( n = 3). The protein bands of WB were derived from different gels, and the uncropped original WB images are provided in the Supplementary Material 1. Data are presented as mean ± SD, differences between groups were analyzed by one-way ANOVA, statistical significance was determined using one-way ANOVA. * P < 0.05, **<0.01, ***<0.001, ****<0.0001 (vs. TGF-β1 group); # P < 0.05, ##<0.01, ###<0.001, ####<0.0001 (vs. YBX1-oe + NC-sh + TGF-β1 group); ^ P < 0.05, ^^<0.01, ^^^<0.001, ^^^^<0.0001 (vs. NC-oe+PPIB-sh + TGF-β1 group). sh: short hairpin, gene expression interference by introducing short hairpin RNA. oe: overexpression, increasing gene expression by introducing the target gene sequence. NC: Negative control, for sh, refers to lentivirus construction by transfecting plasmids with a scrambled shRNA sequence; for oe, refers to lentivirus construction by directly transfecting empty plasmids. ns: not significant.

Article Snippet: The antibodies used included: anti-PPIB antibody (CUSABIO, China; #CSB-PA018472ESR1HU), anti-YBX1 antibody (Abclonal, China; #A3534), anti-VE-cadherin antibody (Abclonal, China; #A12416), anti-CD31 antibody (Abclonal, China; #A19014), anti-α-SMA antibody (Affinity, China, #AF1032), and anti-GAPDH antibody (Abclonal, China; #A19056).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Immunofluorescence, Staining, Wound Healing Assay, Derivative Assay, Gene Expression, shRNA, Over Expression, Sequencing, Negative Control

Journal: Materials Today Bio

Article Title: Modulation of endothelial-to-mesenchymal transition via NRP-1 targeting with melittin attenuates pulmonary fibrosis

doi: 10.1016/j.mtbio.2025.102659

Figure Lengend Snippet: MLT attenuates TGF-β1-induced EndMT by inhibiting the TGF-β/Smad and MAPK pathways in HUVECs. (A–D) The mRNA levels of endothelial (CD31, VE-cadherin) and mesenchymal (α-SMA, vimentin) markers were determined by RT-qPCR. (E) EndMT-related protein expression of CD31, VE-cadherin, α-SMA, vimentin, Twist, and Snail was examined by western blot. The original western blot images are provided in Quantitative analysis of CD31, VE-cadherin, α-SMA, vimentin, Twist, and Snail protein levels (relative to β-actin). (L) Immunofluorescence staining of EndMT markers in HUVECs. Scale bar: 50 μm; Blue: DAPI; Green: CD31; Magenta: VE-cadherin; Red: α-SMA; Orange: vimentin. (M) Nuclear transcription factor immunofluorescence during EndMT in HUVECs. Scale bar: 50 μm; Blue: DAPI; Green: Twist; Red: Snail. (N) Protein levels of key components in the TGF-β/Smad and MAPK pathways were assessed by western blot. The original western blot images are provided in Quantitative analysis of Smad4/β-actin, p-Smad2/3/Smad2/3, p-ERK/ERK, p-JNK/JNK and p-P38/P38 protein levels. Data are presented as the mean ± SEM (n = 3); ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: Antibodies: VE-cadherin, β-actin and HRP-labeled antibodies were purchased from Boster Company.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Immunofluorescence, Staining

Journal: Materials Today Bio

Article Title: Modulation of endothelial-to-mesenchymal transition via NRP-1 targeting with melittin attenuates pulmonary fibrosis

doi: 10.1016/j.mtbio.2025.102659

Figure Lengend Snippet: Nanoparticle delivery of MLT potentiates anti-PF efficacy. (A) Experimental timeline of BLM-induced PF and therapeutic intervention. Mice received intratracheal BLM on day 0, followed by intraperitoneal administration of pLNPs, MLT-L (0.5 mg/kg), MLT-H (2 mg/kg), M-pLNPs (0.5 mg/kg as MLT) every other day from day 7 to day 21. (B) Representative macroscopic morphological images of lung tissues from each treatment group. (C) Histopathological analyses of lung tissues. Scale bar: 20 μm. (D) Quantitative analysis of fibrosis severity by Ashcroft scoring. (E) Quantitative assessment of Masson staining. (F) Representative Micro-CT images of lung tissue at three anatomical planes: sagittal, coronal, and transverse, demonstrating the spatial distribution of fibrotic lesions. Scale bar: 15 mm. (G) Immunofluorescence staining of key therapeutic targets in lung tissues. Scale bar: 50 μm. Blue: DAPI; Red: CD31; Yellow: VE-cadherin; Magenta: α-SMA; Green: COL-I; Cyan: Vimentin. (H) Immunofluorescence staining images showing the expression and localization of nuclear transcription factors Twist and Snail in lung tissues. Scale bar: 50 μm. Blue: DAPI; Red: Twist; Magenta: Snail. (I–N) Quantitative analysis of pulmonary function parameters including respiratory system resistance (Rrs) (I), elastance (Ers) (J), compliance (Crs) (K), forced vital capacity (FVC) (L) and forced expiratory volume at 0.2 s (FEV0.2) (M), and static compliance (Cst) (N). Data are presented as the mean ± SEM (n = 5); ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns: not significant.

Article Snippet: Antibodies: VE-cadherin, β-actin and HRP-labeled antibodies were purchased from Boster Company.

Techniques: Staining, Micro-CT, Immunofluorescence, Biomarker Discovery, Expressing