recombinant human vegf 165  (Sino Biological)

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

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

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

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

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