Journal: Bioactive Materials

Article Title: Targeting VEGFR2 inhibition within a spatially-confined conduit promotes nerve self-resolution and alleviates mechanical allodynia

doi: 10.1016/j.bioactmat.2026.03.009

Figure Lengend Snippet: Efficacy of GelMA MAVP MPs in promoting nerve end interface self-resolution. ( A ) Schematic of the peripheral sciatic nerve ligation (p-SNL) model with four experimental groups (i.e., MAVP, VAN, vehicle, and control) ( B ) Immunofluorescence (IF) staining of p-VEGFR2 and YAP (indicating mechanotransduction signaling). ( C ) The positive area percentage of p-VEGFR2 (n = 6). ( D ) Percentage of YAP in nuclear/cytoplasm (n = 6). ( E ) IF staining of proliferation signal (Ki-67) and vessel signal (CD31) for p-SNL animal. ( F ) Quantification of Ki-67/CD31 co-localization area percentage (n = 6). ( G ) IF co-staining of Ki-67 and macrophage marker F4/80. ( H ) Quantification of Ki-67/F4/80 co-localization area percentage (n = 6). ( I ) IF staining of scar marker α-SMA. ( J ) Quantification of α-SMA-positive area percentage (n = 6). Mean values are shown and error bars represent ± s.d., as analyzed by one-way ANOVA followed by the Tukey-Kramer test in ( C , D , F , H and J ). Biological replicates were used for all experiments. ns, p > 0.05, ∗p < 0.05, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: The following primary antibodies were used for the subsequent steps: anti-Yap (mouse, 1:200, Santa sc-376830); anti-p-VEGFR2 (rabbit, 1:100 Invitrogen, PA5-105765); α-SMA (rabbit, 1:200, Proteintech 14395-1-AP); Reca-1 (mouse, 1:200, Santa sc-52665); anti-CD31 (mouse, 1:200, Santa sc-13537); anti-Ki67 (rabbit, 1:150, Cell Signaling 9129S); anti-NF-200 (mouse, 1:200, Sigma, SAB4200747); anti-MBP (rabbit, 1:200, Abcam ab218011); anti-F4/80 (mouse, 1:200, Santa sc-377009); Iba-1 (rabbit, 1:150, Abcam ab178846); anti-CGRP (rabbit, 1:400, Abcam ab283568); anti-TRPA1 (mouse, 1:200, Santa sc-376495); anti-CD86 (rabbit, 1:200, Proteintech 30691-1-AP); CD206 (rabbit, 1:200, Proteintech 18704-1-AP).

Techniques: Ligation, Control, Immunofluorescence, Staining, Marker

Journal: Bioactive Materials

Article Title: Targeting VEGFR2 inhibition within a spatially-confined conduit promotes nerve self-resolution and alleviates mechanical allodynia

doi: 10.1016/j.bioactmat.2026.03.009

Figure Lengend Snippet: Expression of pain signal proteins in peripheral nerve locations. ( A ) Immunohistochemical (IHC) imaging of VEGFA and ( B ) quantification of VEGFA mean integrated density (n = 6). ( C ) IHC staining for NGF and ( D ) quantification of NGF mean integrated density (n = 6). ( E ) IF staining for macrophages (F4/80) and ( F ) quantification of macrophage number per 10 4 μm 2 (n = 6). ( G ) IF staining for scar tissue (α-SMA) and ( H ) quantification of α-SMA -positive area percentage (n = 6). ( I ) IF staining for myelin sheath (MBP) and axon (NF200) and ( J ) quantification of myelin sheath to axon area ratio (n = 6). ( K ) IF staining for pain-related mediators CGRP and TRPA1 and ( L ) quantification of CGRP (n = 6), and ( M ) TRPA1 (n = 6). Mean values are shown and error bars represent ± s.d., as analyzed by one-way ANOVA followed by the Tukey-Kramer test in ( B , D , F , H , J , L and M ). Biological replicates were used for all experiments. ns, p > 0.05, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: The following primary antibodies were used for the subsequent steps: anti-Yap (mouse, 1:200, Santa sc-376830); anti-p-VEGFR2 (rabbit, 1:100 Invitrogen, PA5-105765); α-SMA (rabbit, 1:200, Proteintech 14395-1-AP); Reca-1 (mouse, 1:200, Santa sc-52665); anti-CD31 (mouse, 1:200, Santa sc-13537); anti-Ki67 (rabbit, 1:150, Cell Signaling 9129S); anti-NF-200 (mouse, 1:200, Sigma, SAB4200747); anti-MBP (rabbit, 1:200, Abcam ab218011); anti-F4/80 (mouse, 1:200, Santa sc-377009); Iba-1 (rabbit, 1:150, Abcam ab178846); anti-CGRP (rabbit, 1:400, Abcam ab283568); anti-TRPA1 (mouse, 1:200, Santa sc-376495); anti-CD86 (rabbit, 1:200, Proteintech 30691-1-AP); CD206 (rabbit, 1:200, Proteintech 18704-1-AP).

Techniques: Expressing, Immunohistochemical staining, Imaging, Immunohistochemistry, Staining

Journal: ACS Omega

Article Title: The N‑Glycoproteomic Landscape of the Lung in Monocrotaline-Induced Pulmonary Arterial Hypertension

doi: 10.1021/acsomega.6c02324

Figure Lengend Snippet: The flowchart of this study and the validation of MCT-induced PAH rat model. A: The flowchart of the study. B: The typical waveform of pulmonary arterial pressure in MCT-induced PAH rats and control (Ctrl) rats. C–D: The mPAP and RVHI of PAH and control rats. E: H&E staining of lung tissues in PAH rats and controls. F–G. The WT% and WA% of PAH and control rats. H: The results of α-SMA (green), CD68 (purple), and DAPI (blue) staining in PAH lungs at ×400 magnification. The data were expressed as mean ± SD ( n = 8); * p < 0.05 vs Ctrl.

Article Snippet: Formalin-fixed, paraffin-embedded lung sections (4 μm) were deparaffinized in xylene, rehydrated through a graded ethanol series (100% to 50%), and subjected to antigen retrieval in EDTA buffer (pH 9.0) at 95 °C for 20 min. After cooling to room temperature and washing with PBS, sections were blocked with 10% nonfat milk for 30 min and then incubated overnight at 4 °C with primary antibodies against α-SMA (1:800; GB13044–50, Servicebio, China), CD68 (1:200; ab318303, Abcam, UK), VCAM1 (1:200, ET1601-18, Huabio, China), and AMBP (1:1000, ER1803-35, Huabio, China).

Techniques: Biomarker Discovery, Control, Staining

Journal: ACS Omega

Article Title: The N‑Glycoproteomic Landscape of the Lung in Monocrotaline-Induced Pulmonary Arterial Hypertension

doi: 10.1021/acsomega.6c02324

Figure Lengend Snippet: Elevation of VCAM1 and AMBP in lungs of MCT-induced PAH rats. A–B: Proteomic and N-glycoproteomic analyses revealed an increase in total protein level and N423-site glycosylation of VCAM1 in MCT-treated rats. C–D: Total protein level and N233-site glycosylation of AMBP were elevated in MCT-treated rats. E: Representative Western blot images showing protein expression of VCAM1 and AMBP in lung tissues. F–G: Quantitative analysis of VCAM1 and AMBP protein levels. H–I: Immunofluorescence results of VCAM1 and AMBP expression in lung tissues. Representative images show α-SMA (red), VCAM1 (green, H) or AMBP (green, I), DAPI (blue), and the merged channels (Merge) at ×400 magnification. Data are presented as mean ± SD; n = 10 (Ctrl), n = 8 (MCT); * p < 0.05, ** p < 0.01, *** p < 0.001 vs Ctrl.

Article Snippet: Formalin-fixed, paraffin-embedded lung sections (4 μm) were deparaffinized in xylene, rehydrated through a graded ethanol series (100% to 50%), and subjected to antigen retrieval in EDTA buffer (pH 9.0) at 95 °C for 20 min. After cooling to room temperature and washing with PBS, sections were blocked with 10% nonfat milk for 30 min and then incubated overnight at 4 °C with primary antibodies against α-SMA (1:800; GB13044–50, Servicebio, China), CD68 (1:200; ab318303, Abcam, UK), VCAM1 (1:200, ET1601-18, Huabio, China), and AMBP (1:1000, ER1803-35, Huabio, China).

Techniques: Glycoproteomics, Western Blot, Expressing, Immunofluorescence

Journal: Brazilian Journal of Cardiovascular Surgery

Article Title: Optimization of the in vitro Model of Cardiac Fibrosis

doi: 10.21470/1678-9741-2025-0075

Figure Lengend Snippet: The expression of alpha-smooth muscle actin (α-SMA) protein. A) Immunostaining of α-SMA. (B): Quantification of α-SMA intensity expression in positive cells. Data are expressed as mean ± standard deviation (three separate experiments with three replicates in each). *P < 0.05. AA=ascorbic acid; Ang II=angiotensin II; Dex=dextran sulfate.

Article Snippet: Next, the cells were incubated with primary antibodies against α-SMA (mouse, Zytomed Systems) or vimentin (1:400, mouse, Sigma, Germany) 75 minutes at room temperature and overnight at 4°C, respectively.

Techniques: Expressing, Immunostaining, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: Partial EMT phenotype proteins in HK-2 cells transfected with lncRNA NKILA overexpression virus (A) Representative western blotting images and (B) quantification of EMT phenotypic indicators. (C) Statistical analysis results of E-cad fluorescence intensity (n=3). (D) RT-qPCR statistical results of EMT phenotype indicators (n=3). ‘Normal’ indicates after starvation treatment, HK-2 cells were replaced with fresh complete medium and continued to be cultured for 24 h. Control, following starvation treatment, 10 ng/ml TGF-β1 cytokine diluent was added to induce HK-2 cells to construct the renal interstitial fibrosis model for 24 h. ‘OE-NKILA’ indicates after starvation, overexpressed Lv-NKILA (76304) was used for transfection and cells were collected after 24 h of lentivirus transfection. Compared with OE-NC ## P<0.01. E-cad; epithelial-cadherin; RT-qPCR, reverse transcription quantitative PCR; EMT, epithelial-mesenchymal transition; NC, negative control; Lv, lentivirus; OE, overexpression; FN, fibronectin; Col1, collagen I; α-SMA, α-smooth muscle actin; Vim; vimentin; ns, not significant.

Article Snippet: After blocking with 5% non-fat milk for 2 h at room temperature, membranes were incubated overnight at 4°C with primary antibodies against fibronectin (FN; 1:1,000; cat. no. ab45688; Abcam), collagen I (Col1; 1:1,000; cat. no. ab138492; Abcam), vimentin (Vim; 1:1,000; cat. no. 10366-1-AP; Proteintech Group, Inc.), α-smooth muscle actin (α-SMA; 1:1,000; cat. no. 14395-1-AP; Proteintech Group, Inc.), JAK2 (1:1,000; cat. no. 17670-1-AP; Proteintech Group, Inc.), STAT3 (1:1,000; cat. no. 10253-2-AP; Proteintech Group, Inc.), phosphorylated (p)-JAK2 (1:500; cat. no. ab32101; Abcam), p-STAT3 (1:500; cat. no. ab76315; Abcam) and GAPDH (1:5,000; cat. no. 10494-1-AP; Proteintech Group, Inc.).

Techniques: Transfection, Over Expression, Virus, Western Blot, Fluorescence, Quantitative RT-PCR, Cell Culture, Control, Construct, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: Partial EMT phenotype proteins in HK-2 cells transfected with lncRNA NKILA knockdown lentivirus (A) Representative western blotting bands and (B) semi-quantification results of EMT phenotypic indicators. (C) E-cad fluorescence intensity and (D) EMT phenotypic index RT-qPCR (n=3). ‘Normal’ indicates starvation treatment. Compared with normal group ## P<0.01. Compared with control + KD-NC group, **P<0.01 and *P<0.05. EMT, epithelial-mesenchymal transition; E-cad, epithelial-cadherin; RT-qPCR, reverse transcription quantitative PCR; KD, knockdown; Lv, lentivirus; shRNA, short hairpin RNA; NC, negative control; FN, fibronectin; Col1, collagen I; α-SMA, α-smooth muscle actin; Vim; vimentin.

Article Snippet: After blocking with 5% non-fat milk for 2 h at room temperature, membranes were incubated overnight at 4°C with primary antibodies against fibronectin (FN; 1:1,000; cat. no. ab45688; Abcam), collagen I (Col1; 1:1,000; cat. no. ab138492; Abcam), vimentin (Vim; 1:1,000; cat. no. 10366-1-AP; Proteintech Group, Inc.), α-smooth muscle actin (α-SMA; 1:1,000; cat. no. 14395-1-AP; Proteintech Group, Inc.), JAK2 (1:1,000; cat. no. 17670-1-AP; Proteintech Group, Inc.), STAT3 (1:1,000; cat. no. 10253-2-AP; Proteintech Group, Inc.), phosphorylated (p)-JAK2 (1:500; cat. no. ab32101; Abcam), p-STAT3 (1:500; cat. no. ab76315; Abcam) and GAPDH (1:5,000; cat. no. 10494-1-AP; Proteintech Group, Inc.).

Techniques: Transfection, Knockdown, Western Blot, Fluorescence, Quantitative RT-PCR, Control, Reverse Transcription, Real-time Polymerase Chain Reaction, shRNA, Negative Control

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: Detection of EMT-related phenotypic proteins in the AG490 intervention HK-2 cell recovery experiment (A) Representative western blotting bands and (B) semi-quantification of EMT phenotypic protein changes (n=3). (C) Statistical analysis results of E-cad fluorescence intensity (n=3) and (D) EMT phenotypic index through reverse transcription quantitative PCR (n=3). ‘Normal’ indicates starvation treatment, HK-2 cells were replaced with fresh complete medium and continued to be cultured for 24 h. ## P<0.01 compared with the normal group, ** P<0.01 and *P<0.05 compared with the control + DMSO group and △△ P<0.01 and △ P<0.05 compared with the OE-NKILA group. EMT, epithelial-mesenchymal transition; E-cad, epithelial cadherin; OE, overexpression; Lv, lentivirus; FN, fibronectin; Col1, collagen I; α-SMA, α-smooth muscle actin; Vim; vimentin.

Article Snippet: After blocking with 5% non-fat milk for 2 h at room temperature, membranes were incubated overnight at 4°C with primary antibodies against fibronectin (FN; 1:1,000; cat. no. ab45688; Abcam), collagen I (Col1; 1:1,000; cat. no. ab138492; Abcam), vimentin (Vim; 1:1,000; cat. no. 10366-1-AP; Proteintech Group, Inc.), α-smooth muscle actin (α-SMA; 1:1,000; cat. no. 14395-1-AP; Proteintech Group, Inc.), JAK2 (1:1,000; cat. no. 17670-1-AP; Proteintech Group, Inc.), STAT3 (1:1,000; cat. no. 10253-2-AP; Proteintech Group, Inc.), phosphorylated (p)-JAK2 (1:500; cat. no. ab32101; Abcam), p-STAT3 (1:500; cat. no. ab76315; Abcam) and GAPDH (1:5,000; cat. no. 10494-1-AP; Proteintech Group, Inc.).

Techniques: Cell Recovery, Western Blot, Fluorescence, Reverse Transcription, Real-time Polymerase Chain Reaction, Cell Culture, Control, Over Expression