Fig 1: Patients with high-circulating PlGF levels, post-NAC, showed a better response. Univariate analysis revealed a significant decrease in plasma PlGF levels in breast cancer patients measured prior to the initiation of chemotherapy (pre-NAC) compared to the levels measured in healthy controls (A). The graphs (B–E) represent the paired t-test results, which show the mean plasma PlGF ± SEM. A significant increase in the PlGF level was detected in patients who were tumor-free after finishing NAC cycles, ypT0 (B), and in patients who responded completely to the treatment, pCR (C). The graphs (D,E) show PlGF levels increased significantly post-NAC in patients aged between 36–50 and 51–70 years, and in patients who have a BMI >30. p ≤ 0.05 is considered to indicate statistical significance.
Fig 2: Maternal LIPUS treatment increased neurotrophic factor expression in IUGR brains. Representative immunoblots and quantitative data of (A) BDNF, (B) PlGF, (C) VEGF, and (D) GDNF at PND 1 after IUGR. Values are presented as mean ±standard error of mean; * p < 0.05 and ** p < 0.01 versus saline group; # p < 0.05 versus DEX group (n = 9–10/group; one‐way analysis of variance). BDNF, brain‐derived neurotrophic factor; DEX, dexamethasone; GDNF, glial cell line‐derived neurotrophic factor; IUGR, intrauterine growth restriction; L, low‐intensity pulsed ultrasound; P1GF, placental growth factor; PND, postnatal day; VEGF, vascular endothelial growth factor
Fig 3: Elevated plasma NRP-1 and PlGF associates with advanced breast cancer. The graphs represent mean concentration of plasma NRP-1 ± SEM measured by ELISA in breast cancer patients which indicates significant upregulation in (a) advanced nodal disease and (b) metastasis, (c) advanced disease stage and (d) tumor size. (e) Plasma PlGF mean concentration ± SEM shows significantly upregulated levels in metastatic breast cancer cases compared to non-metastatic and healthy controls and (f) upregulation in Stage 4 cases compared to stage 2 and 3. p < 0.05 considered to indicate statistical significance. *p < 0.05, **p < 0.01, ***p < 0.001, Multivariate ANOVA followed by Fisher’s LSD post hoc test.
Fig 4: Mode of action of Ate-Grab within the tumor microenvironment (TME).The functional effects of Ate-Grab in the pancreatic tumor microenvironment are briefly summarized and illustrated. We developed a multi-paratopic-VEGF decoy receptor, dubbed “Ate-Grab,” by fusing the single-chain Fv of atezolizumab (an anti-PD-L1 antibody) to the N-terminus of VEGF-Grab. Ate-Grab treatment blocked and sequestered PlGF/VEGF around NRP1+ CAFs (cancer-associated fibroblasts). Simultaneously, Ate-Grab dramatically reduced the number of CD141+ CAFs (“CAF-2”), activating collagen-secreting CAFs. This results in the promotion of cytotoxic T-cell expansion within the TME, thereby suppressing tumor growth compared to the control group.
Fig 5: Inhibitory effect of D16F7 mAb on transmigration of human melanoma cells across an activated endothelial cell monolayer in response to human PlGF. (A) Monolayers of human endothelial cells (HUV-ST cells) were seeded on the upper side of Boyden chambers and activated with TNF-α. Human melanoma WM115 cells were labelled with calcein-AM and incubated for 30 min at room temperature in the absence (UT) or presence of 5 μg/mL D16F7 mAb or mouse IgG as a control (mIgG1). WM115 cells (2 × 105 cells/chamber) were then loaded on the HUV-ST-activated monolayer and allowed to migrate for 5 h against PlGF (50 ng/mL), which was added as stimulus in the lower compartment of the Boyden chamber. Fluorescence of migrated cells was measured in four to eight fields/filter, and results were expressed as relative transmigration compared to unstimulated cells (UNS). Values in the histogram represent the mean ± SD of five Boyden chambers from three independent experiments. Statistical analysis was performed by Kruskal–Wallis test, followed by Dunn’s post hoc analysis: *, p < 0.05; ns, not significant. (B) Representative photographs of fluorescent WM115 cells migrated to the lower side of the filter through the endothelial monolayer are shown for each experimental condition (100× magnification). The scale bars represent 100 µm.
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