Journal: International Journal of Biological Sciences

Article Title: Targeting NCAPG with Arundanine Suppresses Neutrophil/NET-Mediated Immune Evasion and Enhances CD8 + T Cell Function in Lung Squamous Cell Carcinoma

doi: 10.7150/ijbs.134668

Figure Lengend Snippet: The E2F8/NCAPG axis affects TAN phenotype and tumorigenesis in mice. KLN205 cells stably transfected with E2F8 KD and NCAPG OE were implanted into DBA/2 mice subcutaneously. (A) Growth rate of subcutaneous tumors in mice over 28 days. (B) Representative images and weight of in mice on day 28. (C-D) Populations of iNOS + Ly6G + (N1) and Arg1 + Ly6G + (N2) neutrophils in tumor tissues determined using double label immunofluorescence staining. (E) Concentrations of IL-8 and VEGFA in tumors determined using ELISA kits. (F) Levels of p-SMAD2 and p-SMAD3 in tumors determined using WB analysis. (G) Cell apoptosis in tumors determined using TUNEL assay. (H) Expression of E2F8 and NCAPG in tumor tissues analyzed using IHC. (I-K) Flow cytometry detection of the proportion of total CD3⁺CD45⁺CD8⁺ T cells (I), GZMB⁺ (J) or TNF-α⁺ CD8⁺ (K) T cells in tumor tissues. (L) KLN205 mouse lung cancer cells were subjected to E2F8 KD and NCAPG OE transfections and then implanted into DBA/2 mice via tail vein injection, followed by Kaplan-Meier analysis for 70-day survival rate of mice in each group. Data are expressed as the mean ± SD and shown as dot plots with whiskers representing SD. In vitro assays represent n = 6 independent biological replicates. For animal experiments, n = 6 biologically independent mice were analyzed per group, except n = 10 per group in panel L. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The contents of tumor necrosis factor α (TNF-α), interleukin-8 (IL-8), and vascular endothelial growth factor A (VEGFA) in TAN supernatant and tumor tissue homogenates were quantified using ELISA kits for Human IL-8 (E-EL-H6008, Elabscience Biotechnology Co., Ltd.), Human TNF-α (E-EL-H0109, Elabscience), Human VEGFA (E-EL-H0111, Elabscience), Mouse KC (IL-8, MBS8579382, Mybiosource, San Diego, CA, USA), and Mouse VEGFA (E-EL-M1292, Mybiosource).

Techniques: Stable Transfection, Transfection, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Analysis, TUNEL Assay, Expressing, Flow Cytometry, Injection, In Vitro

Journal: Bioactive Materials

Article Title: AdMSC spheroids encapsulating antioxidant hybrid protein carrier for irradiation-damaged salivary gland repair

doi: 10.1016/j.bioactmat.2026.03.049

Figure Lengend Snippet: Functional characterization of AdMSC spheroids and their integration with GC. (a) Bright-field images and size distribution of spheroids generated in agarose molds targeting a diameter of 300 μm. (b) Representative confocal images of hypoxic core staining in spheroids, showing nuclei (blue) and hypoxic regions (green). (c, d) ELISA quantification of cumulative VEGF and HGF secretion from 2D-cultured AdMSCs and spheroids after 3 days culture. (e) Bright-field and confocal fluorescence images of spheroids alone, spheroids exposed to blue light (B.L), spheroids encapsulating 5% (w/v) gelatin hydrogel, or spheroids encapsulating 4–7% (w/v) GCs and photo-crosslinked under 450 nm blue light. Samples were stained with phalloidin (green, F-actin) and DAPI (blue, nuclei); dotted boxes indicate magnified regions. (f) Quantification of normalized cell proliferation within the GC in each condition over 1, 3, and 5 days incubation (n = 3). (g) Cumulative VEGF secretion from encapsulated spheroids measured at 1, 3, and 5 days (n = 5). (h) Spheroid diameter measurements over the incubation period, indicating the degree of spheroid spreading within each matrix (n = 10). (i) Representative HUVEC tube formation images after treatment with conditioned media from VEGF-free control (-VEGF), VEGF-supplemented media (+VEGF), spheroids alone, spheroid encapsulating 5% (w/v) GC, or spheroid encapsulating 7% (w/v) GC groups. (j–m) Quantitative tube formation parameters, including total tube length, number of meshes, number of junctions, and total mesh area, respectively (n = 4). (n) Representative Annexin V-FITC/propidium iodide flow cytometry plots of H 2 O 2 -treated SG cells in the absence or presence of spheroid encapsulating 5% (w/v) GC or spheroid encapsulating 7% (w/v) GCs. (o) Quantification of live, early apoptotic, and late apoptotic cell populations under each condition (n = 3). (p) Relative M2/M1 polarization of macrophages after LPS stimulation and treatment with conditioned media with spheroids and spheroids encapsulating GCs as determined by ELISA (n = 3). Data are presented as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. ns P > 0.05, ∗ P ≤ 0.05, ∗∗ P ≤ 0.01, and ∗∗∗ P ≤ 0.001.

Article Snippet: VEGF and HGF levels were quantified using human VEGF (ABclonal, RK00023) and human HGF ELISA kits according to the manufacturers’ protocols, and absorbance was read at 450 nm with 540 nm reference using a microplate reader (VersaMax; Molecular Devices, USA).

Techniques: Functional Assay, Generated, Staining, Enzyme-linked Immunosorbent Assay, Cell Culture, Fluorescence, Incubation, Control, Flow Cytometry

Journal: Bioactive Materials

Article Title: AdMSC spheroids encapsulating antioxidant hybrid protein carrier for irradiation-damaged salivary gland repair

doi: 10.1016/j.bioactmat.2026.03.049

Figure Lengend Snippet: Angiogenic effect of spheroids encapsulating GC for IR-damaged salivary glands. (a) Immunofluorescence staining for CD31 (green), an endothelial cell marker, in SG tissue sections to evaluate neovascularization. (b, c) Quantification of CD31-positive areas at 1 and 6 weeks post-treatment. (d) Immunofluorescence staining for VEGF-A (red), an angiogenesis marker, in SG tissue sections. (e, f) Quantification of VEGF-A-positive areas at 1 and 6 weeks post-treatment. (g–n) Relative mRNA expression levels of angiogenesis-related genes, including VEGF, HGF, CD31, and vWF, in mouse SG tissues harvested at 1 and 6 weeks post-treatment, as determined by quantitative PCR. Data are presented as mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. ns P > 0.05, ∗ P ≤ 0.05, ∗∗ P ≤ 0.01, and ∗∗∗ P ≤ 0.001.

Article Snippet: VEGF and HGF levels were quantified using human VEGF (ABclonal, RK00023) and human HGF ELISA kits according to the manufacturers’ protocols, and absorbance was read at 450 nm with 540 nm reference using a microplate reader (VersaMax; Molecular Devices, USA).

Techniques: Immunofluorescence, Staining, Marker, Expressing, Real-time Polymerase Chain Reaction

Journal: Nature Communications

Article Title: PCSK5 promotes angiogenesis and cardiac repair after myocardial infarction

doi: 10.1038/s41467-026-72148-7

Figure Lengend Snippet: A The Venn diagram illustrated 104 genes that were commonly upregulated in both infarcted mice ( GSE184986 ) and in infarcted patients ( GSE132143 ). The top 7 secretory proteins were shown. B qPCR analysis detected the PCSK5 expression at the border zone of left ventricular (LV) in both controls and patients with myocardial infarction (MI), n = 5 per group. C ELISA measured plasma PCSK5 levels in controls ( n = 30) and patients with AMI ( n = 28). D Receiver operating characteristic curve was generated for plasma PCSK5 levels to identify AMI patients who exhibited an increased ejection fraction after a six-month follow-up ( n = 28). E Western blot analysis measured the expressions of PCSK5 and GAPDH at the border zone of LV in sham mice and mice post-MI, n = 6 per group. F , G The expressions of PCSK5 and GAPDH were measured by Western blot ( F ) and qPCR ( G ) analyses at the border, infarct, and remote zone of hearts from sham and mice at 28 days post-MI, n = 6 per group. H Immunostaining at the border zone of LV indicated the expressions of PCSK5 and CD31 in both controls ( n = 6) and MI patients ( n = 5). Scale bar represents 60 μm. I Immunostaining at the border zone of LV indicated the expressions of PCSK5 and VE-cadherin (VE-CAD) in sham and mice at 28 days post-MI. Scale bar represents 30 μm, n = 6 per group. J Western blot analysis measured the expressions of PCSK5 and GAPDH in the endothelial cells (EC), cardiomyocytes (CM), and fibroblasts (FB) isolated from the hearts of sham and mice at 28 days post-MI, n = 6 per group. n represents biological replicates. Statistical analyses were performed by two-sided Welch’s t -test for ( B ), two-sided Mann–Whitney U test for ( C and J ) (EC), and by two-sided Student’s t -test for ( H – J ) (CM and FB), and by one-way ANOVA followed by Tukey’s test for ( E and F ), and Kruskal–Wallis test followed by Dunn’s test for ( G ). Data are presented as mean values ± SD. Source data are provided as a Source Data file.

Article Snippet: Plasma VEGFA level was measured using the human VEGFA ELISA Kit (ABclonal, Hubei, China, #RK00023) or mouse VEGFA ELISA Kit (ABclonal, Hubei, China, #RK00028) according to the manufacturer’s instructions.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Generated, Western Blot, Immunostaining, Isolation, MANN-WHITNEY

Journal: Nature Communications

Article Title: PCSK5 promotes angiogenesis and cardiac repair after myocardial infarction

doi: 10.1038/s41467-026-72148-7

Figure Lengend Snippet: A A schematic diagram was created in BioRender. Guo, J. (2026) https://BioRender.com/loszhp6 . It depicted the MI operation in endothelial cells-deficient Pcsk5 ( Pcsk5 ECKO ) mice and wild-type littermates ( Pcsk5 WT ). MI was induced in 8–10 week-old male mice. Echocardiographic assessments were conducted at 0, 7, 14, and 28 days post-surgery. B qPCR analysis detected the Pcsk5 expression in ECs from Pcsk5 WT and Pcsk5 ECKO mice ( n = 6 per group). C ELISA analysis detected the plasma PCSK5 levels in Pcsk5 WT and Pcsk5 ECKO mice, both sham-operated and at 28 days post-MI, n = 6 per group. D Echocardiography showed ejection fraction (EF%) and fractional shortening (FS%) of Pcsk5 ECKO and Pcsk5 WT mice at day 0, 7, 14, and 28 post-MI. n = 3 for Pcsk5 WT + Sham and Pcsk5 ECKO + Sham, n = 7 for Pcsk5 WT + MI and Pcsk5 ECKO + MI. E Masson staining elucidated the infarcted area from suture to apex in Pcsk5 WT and Pcsk5 ECKO mice at 28 days post-MI. n = 6 per group. Scale bar represents 5 mm. F TUNEL staining was performed at the border zone in Pcsk5 WT and Pcsk5 ECKO mice at 28 days post-MI. Scale bar represents 50 μm. n = 6 per group. G A schematic diagram was created in BioRender. Guo, J. (2026) https://BioRender.com/ll1cyb2 . It depicted the MI operation in 6–8-week-old male mice transfected with AAV-EC- Pcsk5 and AAV-Ctrl. Two weeks following the injection, MI was induced. Echocardiographic assessments were at 0, 7, 14, and 28 days post-MI. H qPCR analysis detected the Pcsk5 expression in ECs. I Echocardiography showed EF% and FS% in AAV-EC- Pcsk5 and AAV-Ctrl mice at day 0, 7, 14, and 28 post-MI. J Masson staining elucidated the infarcted area from suture to apex in AAV-EC- Pcsk5 and AAV-Ctrl mice at 28 days post-MI. Scale bar represents 5 mm. K TUNEL staining was performed at the border zone in AAV-EC- Pcsk5 and AAV-Ctrl mice at 28 days post-MI. Scale bar represents 50 μm. n = 6 per group for ( H – K ). n represents biological replicates. Statistical analyses were performed by two-sided Student’s t -test for ( B , E , F , H , J , and K ), and by two-way ANOVA followed by Tukey’s test for ( C , D , and I ). Data are presented as mean values ± SD. Source data are provided as a Source Data file.

Article Snippet: Plasma VEGFA level was measured using the human VEGFA ELISA Kit (ABclonal, Hubei, China, #RK00023) or mouse VEGFA ELISA Kit (ABclonal, Hubei, China, #RK00028) according to the manufacturer’s instructions.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Staining, TUNEL Assay, Transfection, Injection

Journal: Nature Communications

Article Title: PCSK5 promotes angiogenesis and cardiac repair after myocardial infarction

doi: 10.1038/s41467-026-72148-7

Figure Lengend Snippet: A CellChat analysis identified altered signaling pathways in the interactions among all cell types in AAV- Pcsk5 and AAV-Ctrl mice at 7 days post-MI. B Circle plots of significant ( P value ≤ 0.05) cell-cell communication depicted differentially regulated VEGF pathways and interactions in heart tissues of AAV- Pcsk5 and AAV-Ctrl mice at 7 days post-MI. The line thickness indicated the interaction strength of signals from sending and receiving cells. Arrows indicated directionality. (C) Vlnplot analysis displayed the Vegfa expression in ECs from AAV- Pcsk5 and AAV-Ctrl mice at 7 days post-MI. D , E ELISA analysis measured the plasma VEGFA levels in Pcsk5 ECKO and Pcsk5 WT mice, as well as AAV- Pcsk5 and AAV-Ctrl mice at 28 days post-MI, n = 6 per group. F – H The protein expressions of ERK1/2, PCSK5, GAPDH, and the phosphorylation of ERK1/2 at the border zone of LV, were detected by Western blot analysis in AAV- Pcsk5 and AAV-Ctrl mice ( F ) or in AAV-EC- Pcsk5 and AAV-Ctrl mice ( G ) or in Pcsk5 WT and Pcsk5 ECKO mice ( H ) at 28 days post-MI, n = 6 per group. I , J The protein expressions of ERK1/2, PCSK5, and the phosphorylation of ERK1/2 in cell lysate ( I ) or the cleaved VEGFA in cell supernatant ( J ) were detected by Western blot analysis in human umbilical vein endothelial cells (HUVECs) transfected with Ad-Ctrl or Ad-PCSK5. VEGFA cleavage (%) = the cleaved VEGFA/total VEGFA. n = 3 biological replicates per group for ( I ) and n = 4 biological replicates per group for ( J ). K Correlation analyses examined the relationship between plasma levels of PCSK5 and VEGFA in patients with AMI ( n = 42). n represents biological replicates. Statistical analyses were performed by two-sided Welch’s t -test for P-ERK/ERK in ( G ) and PCSK5 in ( F and H ), by two-sided Mann–Whitney U test for P-ERK/ERK in ( F ), and by two-sided Student’s t -test for PCSK5 in ( G and I ), P-ERK/ERK in ( H and I ), and the VEGFA in ( D , E , and J ), two-sided Wilcoxon test for ( C ), permutation tests for ( B ), and by two-sided Spearman’s correlation for ( K ). Data are presented as mean values ± SD. Source data are provided as a Source Data file.

Article Snippet: Plasma VEGFA level was measured using the human VEGFA ELISA Kit (ABclonal, Hubei, China, #RK00023) or mouse VEGFA ELISA Kit (ABclonal, Hubei, China, #RK00028) according to the manufacturer’s instructions.

Techniques: Protein-Protein interactions, Expressing, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Phospho-proteomics, Western Blot, Transfection, MANN-WHITNEY

Journal: Nature Communications

Article Title: PCSK5 promotes angiogenesis and cardiac repair after myocardial infarction

doi: 10.1038/s41467-026-72148-7

Figure Lengend Snippet: A – C Tube formation assays ( A ), EdU assays ( B ), and transwell assays ( C ) were employed to evaluate the tube formation, cell migration, and proliferation in HUVECs transfected with either Ad-Ctrl or Ad-PCSK5, followed by treatment with the VEGFA antibody Bevacizumab (20 ng/ml for 24 h), n = 4 biological replicates per group. Scale bar represents 50 μm. D The interaction mode and binding sites of PCSK5 and VEGFA were predicted by HDOCK analysis. E The Arg158 and Asn164 of PCSK5 were replaced with alanine (designated as PCSK5 mut ). 100 μg/mL recombinant human PCSK5 or PCSK5 mut were incubated with 100 μg/mL human VEGFA peptide at 37 °C for 6 hs. Samples were detected by Western blot analysis ( n = 3 independent experiments). F , G The cleaved VEGFA in cell supernatant ( F ) or the protein expressions of ERK1/2 and PCSK5, and the phosphorylation of ERK1/2 in cell lysate ( G ) were detected by Western blot analysis in HUVECs transfected with Lentivirus-Ctrl (Lenti-Ctrl), Lentivirus-PCSK5 (Lenti-PCSK5), or Lentivirus-PCSK5 mut (Lenti-PCSK5 mut ). VEGFA cleavage (%) = the cleaved VEGFA/total VEGFA. n = 4 biological replicates per group. H The tube formation assays (upper), transwell assays (middle), and EdU assays (bottom) were employed to evaluate the tube formation, cell migration, and proliferation in HUVECs transfected with Lenti-Ctrl, Lenti-PCSK5, or Lenti-PCSK5 mut , n = 4 biological replicates per group. Scale bar represents 50 μm for tube formation and transwell, and 80 μm for EdU. n represents biological replicates. Statistical analyses were performed by two-way ANOVA followed by Tukey’s test for ( A – C ), and by one-way ANOVA followed by Tukey’s test for ( F – H ). Data are presented as mean values ± SD. Source data are provided as a Source Data file.

Article Snippet: Plasma VEGFA level was measured using the human VEGFA ELISA Kit (ABclonal, Hubei, China, #RK00023) or mouse VEGFA ELISA Kit (ABclonal, Hubei, China, #RK00028) according to the manufacturer’s instructions.

Techniques: Migration, Transfection, Binding Assay, Recombinant, Incubation, Western Blot, Phospho-proteomics

Journal: Nature Communications

Article Title: PCSK5 promotes angiogenesis and cardiac repair after myocardial infarction

doi: 10.1038/s41467-026-72148-7

Figure Lengend Snippet: A Transcription factors binding to the PCSK5 promoter (±2000 bp region surrounding the transcription initiation site of PCSK5 ) were predicted by HOMER. A Venn diagram illustrated the differentially expressed transcription factors in MI patients ( GSE132143 ) compared to controls. B HUVECs were transfected with Control or ETS 1 lentivirus, as well as Control or ETS1 siRNAs. RT-qPCR analysis examined mRNA levels of PCSK5 and ETS1 . n = 4 biological replicates. C , D Luciferase activity was detected in HEK293T cells transfected with luciferase reporter constructs containing various segments of the PCSK5 promoter ( C ) or reporter constructs containing PCSK5 promoter and ETS1 binding sites mutation truncations ( D ), followed by transfection with ETS1 -FLAG, n = 6 biological replicates. E Snapshots of ChIP-seq tracks ( GSE41166 ) demonstrated ETS1 enrichment at the promoter of PCSK5 in HUVECs treated with VEGFA (50 ng/ml). F , G The enrichment of ETS1 at the promoter of PCSK5 was validated by ChIP-qPCR analysis in HUVECs treated with VEGFA (100 ng/ml) for 0 or 24 h ( F , n = 5 biological replicates) or in mouse cardiac ECs ( G , n = 4 biological replicates), with KDR serving as a positive control. H , I RT-qPCR analysis examined PCSK5 mRNA levels in HUVECs treated with VEGFA, n = 4 biological replicates. J , K HUVECs were treated with VEGFA (100 ng/ml) for 24 h and transfected with Control or ETS1 siRNAs. The expressions of PCSK5 and ETS1 were detected by RT-qPCR analysis ( J , n = 4 biological replicates) and Western blot analysis ( K , n = 3 biological replicates). L Immunostaining at the border zone of LV indicated the expressions of PCSK5 and ETS1 in both controls ( n = 6) and MI patients ( n = 5). Scale bar represents 60 μm. n represents biological replicates. Statistical analyses were performed by two-sided Welch’s t -test for ( D ) (right) and ( G ), two-sided Mann–Whitney U test for ( B ) (left), ( D ) (left), and ( F ) ( PCSK5 ), by two-sided Student’s t -test for ( B ) (right), ( C and F ) ( KDR ), two-sided Welch ANOVA tests followed by the Dunnett’s T3 test for ( H ), one-way ANOVA followed by Tukey’s test for ( I ) and by two-way ANOVA followed by Tukey’s test for ( J , K ). Data are presented as mean values ± SD. Source data are provided as a Source Data file.

Article Snippet: Plasma VEGFA level was measured using the human VEGFA ELISA Kit (ABclonal, Hubei, China, #RK00023) or mouse VEGFA ELISA Kit (ABclonal, Hubei, China, #RK00028) according to the manufacturer’s instructions.

Techniques: Binding Assay, Transfection, Control, Quantitative RT-PCR, Luciferase, Activity Assay, Construct, Mutagenesis, ChIP-sequencing, ChIP-qPCR, Positive Control, Western Blot, Immunostaining, MANN-WHITNEY

Journal: Nature Communications

Article Title: PCSK5 promotes angiogenesis and cardiac repair after myocardial infarction

doi: 10.1038/s41467-026-72148-7

Figure Lengend Snippet: A A schematic diagram was created in BioRender. Guo, J. (2026) https://BioRender.com/fjpfytl . It depicted the MI operation in Pcsk5 ECKO mice or Pcsk5 WT littermates with semaglutide treatment. Pcsk5 ECKO mice were treated with tamoxifen to conditionally knock out Pcsk5 in endothelial cells. Semaglutide (60 μg/kg) or vehicle was given to mice via subcutaneous injection daily after MI surgery. MI was surgically induced in 8–10 week-old male mice. Echocardiographic assessments were conducted at 0, 7, 14, and 28 days post-surgery. The heart tissues were harvested at 28 days post-MI. B Echocardiography showed EF% and FS% of Pcsk5 ECKO and Pcsk5 WT mice with or without semaglutide treatment at day 0, 7, 14, and 28 post-MI, n = 6 per group. C IB4 and α-SMA staining at the border zone of LV showed the capillary and arteriole densities in Pcsk5 ECKO and wild-type mice with or without semaglutide treatment at 28 days post-MI. Scale bar represents 100 μm. n = 6 per group. D The schematic diagram was created in BioRender. Guo, J. (2026) https://BioRender.com/0w6zmr8 . It illustrated that plasma PCSK5 levels were elevated in AMI patients compared to controls. VEGFA stimulation facilitated the enrichment of ETS1 at the promoter of PCSK5, thereby elevating its expression. The accumulation of PCSK5 facilitated the proteolytic processing of VEGFA, thereby activating the VEGFA signaling pathway and subsequently angiogenesis. EC-specific deficiency of Pcsk5 in mice resulted in exacerbated cardiac dysfunction and attenuated angiogenesis post-MI. Plasma PCSK5 levels could effectively differentiate the improvement in cardiac function among these patients after a 6-month follow-up. Plasma levels of PCSK5 were positively correlated with VEGFA and GLP-1 in patients with AMI. Semaglutide was observed to enhance the expression of PCSK5 via ETS1, thereby augmenting angiogenic activity in ECs. Semaglutide promoted angiogenesis partially by PCSK5 post-MI in mice. n represents biological replicates. Statistical analyses were performed by two-way ANOVA followed by Tukey’s test for ( B and C ). Data are presented as mean values ± SD. Source data are provided as a Source Data file.

Article Snippet: Plasma VEGFA level was measured using the human VEGFA ELISA Kit (ABclonal, Hubei, China, #RK00023) or mouse VEGFA ELISA Kit (ABclonal, Hubei, China, #RK00028) according to the manufacturer’s instructions.

Techniques: Knock-Out, Injection, Staining, Clinical Proteomics, Expressing, Activity Assay