Fig 1: VEGFA signalling is involved in LINC02321-regulated bladder cancer progression. (A) Gene set enrichment analysis demonstrated that the expression of LINC02321 in The Cancer Genome Atlas bladder cancer dataset was associated with VEGFA signalling. (B) Pearson's correlation analysis demonstrated that LINC02321 expression correlated positively with VEGFA expression. (C) Immunohistochemical staining images from the Human Protein Atlas database demonstrated that the intensity of VEGFA staining in bladder cancer tissue was stronger compared with that in adjacent mucosa. LINC02321 regulated mRNA and protein expression levels of VEGFA, demonstrated using reverse transcription-quantitative PCR following transfection with (D) si-LINC02321 (*P<0.05 and ***P<0.001 vs. si-NC) and (E) OE-LINC02321 (***P<0.001 vs. vector) and (F) western blotting, respectively. (G) Survival time of patients with high LINC02321 expression levels was significantly shorter compared with patients with low LINC02321 expression levels. Data are presented as the mean ± standard deviation from three independent experiments. KEGG, Kyoto Encyclopedia of Genes and Genomes; LINC, long intergenic non-coding RNA; NC, negative control; NES, normalized enrichment score; OE, overexpression; si, small interfering RNA; TPM, transcripts per million.
Fig 2: LPS promoted the release of growth factors and cytokines from microglia, and activated the PI3K-AKT pathway of microglia. (A,B) 24 h after administration with LPS, VEGF, FGF2, IL-6, and MMP9 were elevated in the supernatant of HMC3 cells (n = 4). (*P < 0.05, **P < 0.01, ***P < 0.001). (C,D) 24 h after administration with LPS, western blot analysis of PI3K, AKT and P-AKT level in HMC3 cells. ACTIN was detected as the loading control (n = 5). (*P < 0.05, **P < 0.01, ***P < 0.001). (E) 24 h after administration with LPS, western blot analysis of VEGFA, FGF2, HGFα, MMP9 and PDGFβ level in HMC3 cells. ACTIN was detected as the loading control (n = 4). (*P < 0.05, **P < 0.01, ***P < 0.001).
Fig 3: LINC02913 expression affected ADSC cell phenotype under hypoxia and normoxia. A LINC02913 expression in ADSCs cells treated with different oxygen extensions was detected using qRT-PCR; B the transfection efficiency of LINC02913 overexpression plasmid (LINC02913 OE) was verified using qRT-PCR; C the changes in cell viability after LINC02913 overexpression under hypoxia and normoxic conditions were detected using CCK-8 assay; D the changes in the adipogenic ability of cells after LINC02913 overexpression under hypoxia and normoxic conditions were detected using oil red O staining; E after the induction for endothelial differentiation of ADSCs, cell tube formation after LINC02913 overexpression under hypoxia and normoxic conditions were detected using the tube formation assay; F the expression of endothelial cell marker CD31 was detected using IF staining after LINC02913 overexpression under hypoxia and normoxic conditions; G the level of endothelial cell markers (CD133, CD31, VEGFA, and eNOS) after LINC02913 overexpression under hypoxia and normoxic conditions was detected using western blot. **p < 0.01 compared to Nypoxia + vector group; ##p < 0.01, compared with Hypoxia+vector group
Fig 4: Sirt3 deficiency severely impairs VEGFA expression and further reduces glomerular VEGFA upon ADR treatment.(A) Representative Western Blot and quantification of VEGFA in WT and Sirt3-/- mice, treated with saline or ADR (n = 4 mice per groups). (B) Representative images and quantification of glomerular VEGFA in WT and Sirt3-/- mice after 7 weeks receiving saline or ADR (n = 3 mice for group). Scale bars, 20 μm. (C) Representative images of double staining for VEGFA (red) and nestin (green) in WT and Sirt3-/- mice. Scale bars, 10 μm. Data represent mean ± SEM and were analyzed by one-way ANOVA followed by Tukey’s multiple comparisons test. °P<0.05, and °°°P<0.001 vs WT+saline; ###P<0.001 vs WT+ADR.
Fig 5: Efficacy of targeting oncomiRs 10b and 21 via BNP-delivered sγPNAs in a PDX mice model of GBM.(A) Survival of mice-bearing patient (G22)–derived intracranial gliomas after treatment with BNPs containing sγPNA, TMZ, and the combination treatment of BNP with TMZ. BNPs were administered by CED at a dose of 200 mg/ml, and TMZ was administered intraperitoneally at a dose of 25 mg/kg. (B) The levels of miR-21 and miR-10b in gliomas of control and BNP + TMZ–treated mice at the end of the survival study. Results are represented as means ± SD (n = 3 biologically independent animals). The control tumor tissues were harvested after animal euthanasia, and treated mice brain tissues were harvested on day 120. (C) Histology of H&E-stained control and BNP + TMZ–treated mice brain at the end of the survival study. Control mouse brain was harvested on day 39, and sγPNA/BNP + TMZ–treated mouse brain was harvested on day 120. Scale bars, 75 μm. (D) Ki67 staining of control and BNP + TMZ–treated mice brain at the end of the survival study. Control mouse brain was harvested on day 39, and sγPNA/BNP + TMZ–treated mouse brain was harvested on day 120. Scale bars, 75 μm. (E) The expression levels of miR-10b and miR-21 in mice gliomas 48 hours after CED of NPs on day 14 and (F) day 28 after tumor implantation. Results are represented as means ± SEM (n = 3 biologically independent animals). (G) The levels of downstream genes PDGFRA, PDGFRB, VEGFA, ITGB8, ITGA10, ITGA11, ANGPT2, ANGPT1, PRKCA, and IL6 in mice gliomas 48 hours after CED of NPs on day 28. Results are represented as means ± SEM (n = 3 biologically independent animals). sγPNA/BNPs are a physical mixture of sγPNA-21 BNP and sγPNA-10b BNP. NNP indicates PLA-HPG NP, and BNP indicates PLA-HPG-CHO NP.
from Cell Signaling Technology for VEGF-A (E9X8Q) Rabbit mAb