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Scheme 1. General design of P-Nb 2 C-PEG@si-circPUM1 in ovarian cancer. (A) Schematic illustration of P-Nb 2 C-PEG preparation. The precursor (Nb 2 AlC) underwent HF etching followed by exfoliation with TPAOH, yielding negatively charged monolayer Nb 2 C nanosheets. After mixture with excess PEI, P-Nb 2 C was generated with a positive surface charge. We modified the nanosheets with CHO-PEG-CHO, where the aldehyde groups form Schiff base linkages with amine groups, enabling PEG grafting. (B) P-Nb 2 C-PEG was loaded with circPUM1 siRNA via electrostatic interaction and circulated in blood (pH 7.4). Upon reaching the acidic tumor microenvironment (pH 6.5), the nanosheets underwent pH-responsive charge reversal enabling effective cellular uptake. (C) The internalized circPUM1 siRNA specifically targeted the back-splice junction of circPUM1, disrupted its circular structure and abolished its miRNA-sponging activity. The released miRNAs suppressed the expression of <t>VEGFA</t> <t>and</t> <t>RAB27B,</t> thereby inhibiting RAB27B-mediated exosome secretion and VEGFA-triggered angiogenesis. (Created in BioRender. Guan, X. (2025) https://BioRender.com/x7gxaqp )
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MS4A1 inhibits angiogenesis to regulate macrophage M1 infiltration. A The scatter and volcano plot showing the upregulated and downregulated genes in H1650 shNC and shMS4A1 group based on transcriptome sequencing. B The GO functional enrichment analysis based on transcriptome sequencing showing that angiogenesis-related pathways enriched differentially in shNC and shMS4A1 group. C WB experiments indicating the relationship of MS4A1 with <t>VEGFA</t> protein. D CCK-8 experiments indicating the effects of MS4A1 on the proliferation of HUVECs. E Transwell experiments indicating the effects of MS4A1 on the migrated capacity of HUVECs. F , H The tube formation assay indicating the effect of MS4A1 expression on angiogenesis. G IHC analysis demonstrating expression levels of CD31, CD68, and CD80 in tissue samples with high and low MS4A1 expression.
Anti Vascular Endothelial Growth Factor A Vegfa, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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MS4A1 inhibits angiogenesis to regulate macrophage M1 infiltration. A The scatter and volcano plot showing the upregulated and downregulated genes in H1650 shNC and shMS4A1 group based on transcriptome sequencing. B The GO functional enrichment analysis based on transcriptome sequencing showing that angiogenesis-related pathways enriched differentially in shNC and shMS4A1 group. C WB experiments indicating the relationship of MS4A1 with <t>VEGFA</t> protein. D CCK-8 experiments indicating the effects of MS4A1 on the proliferation of HUVECs. E Transwell experiments indicating the effects of MS4A1 on the migrated capacity of HUVECs. F , H The tube formation assay indicating the effect of MS4A1 expression on angiogenesis. G IHC analysis demonstrating expression levels of CD31, CD68, and CD80 in tissue samples with high and low MS4A1 expression.
Vascular Endothelial Growth Factor A Vegfa, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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MS4A1 inhibits angiogenesis to regulate macrophage M1 infiltration. A The scatter and volcano plot showing the upregulated and downregulated genes in H1650 shNC and shMS4A1 group based on transcriptome sequencing. B The GO functional enrichment analysis based on transcriptome sequencing showing that angiogenesis-related pathways enriched differentially in shNC and shMS4A1 group. C WB experiments indicating the relationship of MS4A1 with <t>VEGFA</t> protein. D CCK-8 experiments indicating the effects of MS4A1 on the proliferation of HUVECs. E Transwell experiments indicating the effects of MS4A1 on the migrated capacity of HUVECs. F , H The tube formation assay indicating the effect of MS4A1 expression on angiogenesis. G IHC analysis demonstrating expression levels of CD31, CD68, and CD80 in tissue samples with high and low MS4A1 expression.
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MS4A1 inhibits angiogenesis to regulate macrophage M1 infiltration. A The scatter and volcano plot showing the upregulated and downregulated genes in H1650 shNC and shMS4A1 group based on transcriptome sequencing. B The GO functional enrichment analysis based on transcriptome sequencing showing that angiogenesis-related pathways enriched differentially in shNC and shMS4A1 group. C WB experiments indicating the relationship of MS4A1 with <t>VEGFA</t> protein. D CCK-8 experiments indicating the effects of MS4A1 on the proliferation of HUVECs. E Transwell experiments indicating the effects of MS4A1 on the migrated capacity of HUVECs. F , H The tube formation assay indicating the effect of MS4A1 expression on angiogenesis. G IHC analysis demonstrating expression levels of CD31, CD68, and CD80 in tissue samples with high and low MS4A1 expression.
Vascular Endothelial Growth Factor Vegf, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Proteintech anti-vascular endothelial growth factor a (vegfa) rabbit antibody
MS4A1 inhibits angiogenesis to regulate macrophage M1 infiltration. A The scatter and volcano plot showing the upregulated and downregulated genes in H1650 shNC and shMS4A1 group based on transcriptome sequencing. B The GO functional enrichment analysis based on transcriptome sequencing showing that angiogenesis-related pathways enriched differentially in shNC and shMS4A1 group. C WB experiments indicating the relationship of MS4A1 with <t>VEGFA</t> protein. D CCK-8 experiments indicating the effects of MS4A1 on the proliferation of HUVECs. E Transwell experiments indicating the effects of MS4A1 on the migrated capacity of HUVECs. F , H The tube formation assay indicating the effect of MS4A1 expression on angiogenesis. G IHC analysis demonstrating expression levels of CD31, CD68, and CD80 in tissue samples with high and low MS4A1 expression.
Anti Vascular Endothelial Growth Factor A (Vegfa) Rabbit Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Scheme 1. General design of P-Nb 2 C-PEG@si-circPUM1 in ovarian cancer. (A) Schematic illustration of P-Nb 2 C-PEG preparation. The precursor (Nb 2 AlC) underwent HF etching followed by exfoliation with TPAOH, yielding negatively charged monolayer Nb 2 C nanosheets. After mixture with excess PEI, P-Nb 2 C was generated with a positive surface charge. We modified the nanosheets with CHO-PEG-CHO, where the aldehyde groups form Schiff base linkages with amine groups, enabling PEG grafting. (B) P-Nb 2 C-PEG was loaded with circPUM1 siRNA via electrostatic interaction and circulated in blood (pH 7.4). Upon reaching the acidic tumor microenvironment (pH 6.5), the nanosheets underwent pH-responsive charge reversal enabling effective cellular uptake. (C) The internalized circPUM1 siRNA specifically targeted the back-splice junction of circPUM1, disrupted its circular structure and abolished its miRNA-sponging activity. The released miRNAs suppressed the expression of VEGFA and RAB27B, thereby inhibiting RAB27B-mediated exosome secretion and VEGFA-triggered angiogenesis. (Created in BioRender. Guan, X. (2025) https://BioRender.com/x7gxaqp )

Journal: Materials Today Bio

Article Title: pH-responsive 2D niobium carbide nanosheets for targeted circPUM1 siRNA delivery in ovarian cancer therapy

doi: 10.1016/j.mtbio.2025.102314

Figure Lengend Snippet: Scheme 1. General design of P-Nb 2 C-PEG@si-circPUM1 in ovarian cancer. (A) Schematic illustration of P-Nb 2 C-PEG preparation. The precursor (Nb 2 AlC) underwent HF etching followed by exfoliation with TPAOH, yielding negatively charged monolayer Nb 2 C nanosheets. After mixture with excess PEI, P-Nb 2 C was generated with a positive surface charge. We modified the nanosheets with CHO-PEG-CHO, where the aldehyde groups form Schiff base linkages with amine groups, enabling PEG grafting. (B) P-Nb 2 C-PEG was loaded with circPUM1 siRNA via electrostatic interaction and circulated in blood (pH 7.4). Upon reaching the acidic tumor microenvironment (pH 6.5), the nanosheets underwent pH-responsive charge reversal enabling effective cellular uptake. (C) The internalized circPUM1 siRNA specifically targeted the back-splice junction of circPUM1, disrupted its circular structure and abolished its miRNA-sponging activity. The released miRNAs suppressed the expression of VEGFA and RAB27B, thereby inhibiting RAB27B-mediated exosome secretion and VEGFA-triggered angiogenesis. (Created in BioRender. Guan, X. (2025) https://BioRender.com/x7gxaqp )

Article Snippet: Membranes were subsequently blocked with NcmBlot blocking buffer (NCM Biotech, China) for 10 min. Primary antibody incubation was carried out at 4 °C overnight with specific antibodies against VEGFA (1:1000; Boster, China), RAB27B (1:1000; Boster, China), and β-actin (1:8000; Abbkine, China).

Techniques: Generated, Modification, Activity Assay, Expressing

CircPUM1 promotes angiogenesis in ovarian cancer as the form of exosomes. (A) The expression of circPUM1 was positively related with MVD in ovarian cancer tissue. (B) Experimental design schematic (Created in BioRender. Guan, X. (2025) https://BioRender.com/f5br0q0 ): HUVEC were co-cultured with exosomes derived from circPUM1-overexpressing and control OVCAR3 cells. (C) RT-qPCR showed that circPUM1 was highly expressed in HUVEC after co-cultured with exosomal circPUM1. Exosomal circPUM1 enhanced migration ability (D, E), viability (F), invasiveness (G), and tube formation ability (H) of HUVECs. (I) Circular RNA pull-down assays using biotinylated circPUM1 probes confirmed co-enrichment of circPUM1 and miR-607. (J) Dual-luciferase reporter assays showed miR-607 targeting 3′-UTR in both VEGFA and RAB27B . (K) Western blot revealed that overexpression of circPUM1 upregulated VEGFA and RAB27B , whereas miR-607 downregulated these targets in OVCAR3 cells. Exosomal circPUM1 elevated intracellular VEGFA expression in HUVECs. (L) Molecular schematic (Created in BioRender. Guan, X. (2025) https://BioRender.com/5eldsz5 ): In ovarian cancer, the highly expressed circPUM1 sponges miR-607, upregulating expression of VEGFA and RAB27B . This dual regulation establishes a pro-tumorigenic cascade through two distinct pathways: (1) RAB27B -mediated secretion of exosomal circPUM1, and (2) VEGFA -induced activation of angiogenic signaling.

Journal: Materials Today Bio

Article Title: pH-responsive 2D niobium carbide nanosheets for targeted circPUM1 siRNA delivery in ovarian cancer therapy

doi: 10.1016/j.mtbio.2025.102314

Figure Lengend Snippet: CircPUM1 promotes angiogenesis in ovarian cancer as the form of exosomes. (A) The expression of circPUM1 was positively related with MVD in ovarian cancer tissue. (B) Experimental design schematic (Created in BioRender. Guan, X. (2025) https://BioRender.com/f5br0q0 ): HUVEC were co-cultured with exosomes derived from circPUM1-overexpressing and control OVCAR3 cells. (C) RT-qPCR showed that circPUM1 was highly expressed in HUVEC after co-cultured with exosomal circPUM1. Exosomal circPUM1 enhanced migration ability (D, E), viability (F), invasiveness (G), and tube formation ability (H) of HUVECs. (I) Circular RNA pull-down assays using biotinylated circPUM1 probes confirmed co-enrichment of circPUM1 and miR-607. (J) Dual-luciferase reporter assays showed miR-607 targeting 3′-UTR in both VEGFA and RAB27B . (K) Western blot revealed that overexpression of circPUM1 upregulated VEGFA and RAB27B , whereas miR-607 downregulated these targets in OVCAR3 cells. Exosomal circPUM1 elevated intracellular VEGFA expression in HUVECs. (L) Molecular schematic (Created in BioRender. Guan, X. (2025) https://BioRender.com/5eldsz5 ): In ovarian cancer, the highly expressed circPUM1 sponges miR-607, upregulating expression of VEGFA and RAB27B . This dual regulation establishes a pro-tumorigenic cascade through two distinct pathways: (1) RAB27B -mediated secretion of exosomal circPUM1, and (2) VEGFA -induced activation of angiogenic signaling.

Article Snippet: Membranes were subsequently blocked with NcmBlot blocking buffer (NCM Biotech, China) for 10 min. Primary antibody incubation was carried out at 4 °C overnight with specific antibodies against VEGFA (1:1000; Boster, China), RAB27B (1:1000; Boster, China), and β-actin (1:8000; Abbkine, China).

Techniques: Expressing, Cell Culture, Derivative Assay, Control, Quantitative RT-PCR, Migration, Luciferase, Western Blot, Over Expression, Activation Assay

In vitro antitumor performance of P-Nb 2 C@si-circPUM1 in ovarian cancer cells. (A) Confocal microscopy images showed intracellular uptake of Cy3-labeled circPUM1 siRNA. Cell apoptosis assays (B), Transwell assays (C), wound healing assays (E) and EdU proliferation assays (F) showed that P-Nb 2 C-loaded circPUM1 siRNA effectively suppressed ovarian cancer cell phenotypes. (G) RT-qPCR demonstrated a significant downregulation of circPUM1 expression in lipo2000-transfected and P-Nb 2 C-loaded siRNA groups. Immunofluorescence (D) and Western blot (H) validated the downregulation of VEGFA and RAB27B expression in cells treated with P-Nb 2 C@si-circPUM1. (I) CCK8 assays showed inhibition of ovarian cancer cell viability in lipo2000-transfected and P-Nb 2 C-loaded siRNA groups.

Journal: Materials Today Bio

Article Title: pH-responsive 2D niobium carbide nanosheets for targeted circPUM1 siRNA delivery in ovarian cancer therapy

doi: 10.1016/j.mtbio.2025.102314

Figure Lengend Snippet: In vitro antitumor performance of P-Nb 2 C@si-circPUM1 in ovarian cancer cells. (A) Confocal microscopy images showed intracellular uptake of Cy3-labeled circPUM1 siRNA. Cell apoptosis assays (B), Transwell assays (C), wound healing assays (E) and EdU proliferation assays (F) showed that P-Nb 2 C-loaded circPUM1 siRNA effectively suppressed ovarian cancer cell phenotypes. (G) RT-qPCR demonstrated a significant downregulation of circPUM1 expression in lipo2000-transfected and P-Nb 2 C-loaded siRNA groups. Immunofluorescence (D) and Western blot (H) validated the downregulation of VEGFA and RAB27B expression in cells treated with P-Nb 2 C@si-circPUM1. (I) CCK8 assays showed inhibition of ovarian cancer cell viability in lipo2000-transfected and P-Nb 2 C-loaded siRNA groups.

Article Snippet: Membranes were subsequently blocked with NcmBlot blocking buffer (NCM Biotech, China) for 10 min. Primary antibody incubation was carried out at 4 °C overnight with specific antibodies against VEGFA (1:1000; Boster, China), RAB27B (1:1000; Boster, China), and β-actin (1:8000; Abbkine, China).

Techniques: In Vitro, Confocal Microscopy, Labeling, Quantitative RT-PCR, Expressing, Transfection, Immunofluorescence, Western Blot, Inhibition

In vitro anti-angiogenic performance of P-Nb 2 C@si-circPUM1 in HUVECs. (A) QPCR showed a reduction of circPUM1 levels in exosomes derived from P-Nb 2 C@si-circPUM1-treated ovarian cancer cells. (B) Intracellular circPUM1 level in HUVEC treated with Exo-P-Nb 2 C@si-circPUM1 were markedly lower than those in Exo-control and Exo-P-Nb 2 C@si-scramble groups. CCK-8 and EdU assays demonstrated that Exo-P-Nb 2 C@si-circPUM1 suppressed cell viability (C) and DNA replication capacity (D, E) in HUVECs. Wound healing, Transwell assay and tube formation assay indicated that Exo-P-Nb 2 C@si-circPUM1 treatment significantly attenuated HUVEC migration (F), invasion (G), and angiogenic potential (I) compared to control groups. Immunofluorescence (H, J) and Western blot (K) confirmed downregulation of VEGFA in Exo-P-Nb 2 C@si-circPUM1 treated HUVECs.

Journal: Materials Today Bio

Article Title: pH-responsive 2D niobium carbide nanosheets for targeted circPUM1 siRNA delivery in ovarian cancer therapy

doi: 10.1016/j.mtbio.2025.102314

Figure Lengend Snippet: In vitro anti-angiogenic performance of P-Nb 2 C@si-circPUM1 in HUVECs. (A) QPCR showed a reduction of circPUM1 levels in exosomes derived from P-Nb 2 C@si-circPUM1-treated ovarian cancer cells. (B) Intracellular circPUM1 level in HUVEC treated with Exo-P-Nb 2 C@si-circPUM1 were markedly lower than those in Exo-control and Exo-P-Nb 2 C@si-scramble groups. CCK-8 and EdU assays demonstrated that Exo-P-Nb 2 C@si-circPUM1 suppressed cell viability (C) and DNA replication capacity (D, E) in HUVECs. Wound healing, Transwell assay and tube formation assay indicated that Exo-P-Nb 2 C@si-circPUM1 treatment significantly attenuated HUVEC migration (F), invasion (G), and angiogenic potential (I) compared to control groups. Immunofluorescence (H, J) and Western blot (K) confirmed downregulation of VEGFA in Exo-P-Nb 2 C@si-circPUM1 treated HUVECs.

Article Snippet: Membranes were subsequently blocked with NcmBlot blocking buffer (NCM Biotech, China) for 10 min. Primary antibody incubation was carried out at 4 °C overnight with specific antibodies against VEGFA (1:1000; Boster, China), RAB27B (1:1000; Boster, China), and β-actin (1:8000; Abbkine, China).

Techniques: In Vitro, Derivative Assay, Control, CCK-8 Assay, Transwell Assay, Tube Formation Assay, Migration, Immunofluorescence, Western Blot

Molecular mechanism validation of P-Nb 2 C-PEG@si-circPUM1 anti-angiogenic performance in vivo . (A) H&E staining showed increased necrotic loci within tumors of P-Nb 2 C-PEG@si-circPUM1 group. (B, D) IHC staining of Ki-67 demonstrated a clear reduction in Ki-67-positive cells in P-Nb 2 C-PEG@si-circPUM1 group. (C, F) IHC staining of CD31 indicated a marked decrease in MVD within tumors of P-Nb 2 C-PEG@si-circPUM1 group. (H) QPCR showed a significant downregulation of circPUM1 expression in the P-Nb 2 C-PEG@si-circPUM1 treated group. IHC (E, G) and Western blot (I, J) demonstrated significant down-regulation of RAB27B and VEGFA .

Journal: Materials Today Bio

Article Title: pH-responsive 2D niobium carbide nanosheets for targeted circPUM1 siRNA delivery in ovarian cancer therapy

doi: 10.1016/j.mtbio.2025.102314

Figure Lengend Snippet: Molecular mechanism validation of P-Nb 2 C-PEG@si-circPUM1 anti-angiogenic performance in vivo . (A) H&E staining showed increased necrotic loci within tumors of P-Nb 2 C-PEG@si-circPUM1 group. (B, D) IHC staining of Ki-67 demonstrated a clear reduction in Ki-67-positive cells in P-Nb 2 C-PEG@si-circPUM1 group. (C, F) IHC staining of CD31 indicated a marked decrease in MVD within tumors of P-Nb 2 C-PEG@si-circPUM1 group. (H) QPCR showed a significant downregulation of circPUM1 expression in the P-Nb 2 C-PEG@si-circPUM1 treated group. IHC (E, G) and Western blot (I, J) demonstrated significant down-regulation of RAB27B and VEGFA .

Article Snippet: Membranes were subsequently blocked with NcmBlot blocking buffer (NCM Biotech, China) for 10 min. Primary antibody incubation was carried out at 4 °C overnight with specific antibodies against VEGFA (1:1000; Boster, China), RAB27B (1:1000; Boster, China), and β-actin (1:8000; Abbkine, China).

Techniques: Biomarker Discovery, In Vivo, Staining, Immunohistochemistry, Expressing, Western Blot

MS4A1 inhibits angiogenesis to regulate macrophage M1 infiltration. A The scatter and volcano plot showing the upregulated and downregulated genes in H1650 shNC and shMS4A1 group based on transcriptome sequencing. B The GO functional enrichment analysis based on transcriptome sequencing showing that angiogenesis-related pathways enriched differentially in shNC and shMS4A1 group. C WB experiments indicating the relationship of MS4A1 with VEGFA protein. D CCK-8 experiments indicating the effects of MS4A1 on the proliferation of HUVECs. E Transwell experiments indicating the effects of MS4A1 on the migrated capacity of HUVECs. F , H The tube formation assay indicating the effect of MS4A1 expression on angiogenesis. G IHC analysis demonstrating expression levels of CD31, CD68, and CD80 in tissue samples with high and low MS4A1 expression.

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: MS4A1 regulates M1-polarized tumor-associated macrophage infiltration, angiogenesis, and cancer progression through the HIPPO pathway in lung adenocarcinoma

doi: 10.1007/s00262-025-04213-x

Figure Lengend Snippet: MS4A1 inhibits angiogenesis to regulate macrophage M1 infiltration. A The scatter and volcano plot showing the upregulated and downregulated genes in H1650 shNC and shMS4A1 group based on transcriptome sequencing. B The GO functional enrichment analysis based on transcriptome sequencing showing that angiogenesis-related pathways enriched differentially in shNC and shMS4A1 group. C WB experiments indicating the relationship of MS4A1 with VEGFA protein. D CCK-8 experiments indicating the effects of MS4A1 on the proliferation of HUVECs. E Transwell experiments indicating the effects of MS4A1 on the migrated capacity of HUVECs. F , H The tube formation assay indicating the effect of MS4A1 expression on angiogenesis. G IHC analysis demonstrating expression levels of CD31, CD68, and CD80 in tissue samples with high and low MS4A1 expression.

Article Snippet: Primary antibodies included anti-MS4A1 (1:1000, Proteintech, 60271-1-Ig), anti-vascular endothelial growth factor A (VEGFA) (1:1000, Proteintech, 19003-1-AP), anti-YAP (1:1000, baijia, IPB0668), anti-Phosphorylation YAP (1:1000, baijia, IPH0211), anti-MST (1:1000, Proteintech, 22245-1-AP), anti-Phosphorylation MST (1:1000, Proteintech, 80093-1-RR), anti-TEAD (1:500, Abways, CY9838), and Beta tubulin (1:1000, Proteintech, 66240-1-Ig).

Techniques: Sequencing, Functional Assay, CCK-8 Assay, Tube Formation Assay, Expressing