Journal: Biomedicines

Article Title: Ponciri Fructus Immatarus Sensitizes the Apoptotic Effect of Hyperthermia Treatment in AGS Gastric Cancer Cells through ROS-Dependent HSP Suppression

doi: 10.3390/biomedicines11020405

Figure Lengend Snippet: Effect of co-treatment with PF and hyperthermia on apoptosis in AGS cells. PF was applied to AGS cells (0.3 × 10⁶ cells) for 24 h, with or without hyperthermia. Equal volumes of lysates from whole-cell extracts were then subjected to Western blot analysis. Protein expression levels of ( a ) caspase-3, caspase-8, caspase-9 Bcl-2, Bcl-xL, survivin, ( b ) VEGF, MMP-9, and MMP-2 were measured using Western blot assays. β-actin was used as a loading control. ( c ) Annexin V staining was performed to detect apoptotic cells by flow cytometry. MMP-2, matrix metallopeptidase-2; MMP-9, matrix metallopeptidase-9; PF, Ponciri Fructus Immaturus; VEGF, vascular endothelial growth factor.

Article Snippet: After blocking, the matched primary antibodies (1:3000) were applied to the membranes, including the following primary antibodies at 4 °C overnight: anti-caspase-3, anti-survivin, anti-heat shock protein (HSP) 27, anti-HSP70, anti-HSP90, anti-caspase-8, anti-caspase-9, anti-p-extracellular signal-regulated kinase (ERK) (Thr202/Tyr204), anti-ERK, anti-p-Jun N-terminal kinase (JNK) (Thr183/Tyr185), anti-JNK, anti-p-p38 (Thr180/Tyr182), anti-p38 (Cell Signaling Technology), anti-β-actin, anti-Bcl-2, anti-Bcl-xL, anti-cyclin D1, anti-vascular endothelial growth factor (VEGF), anti-matrix metallopeptidase (MMP) 9, anti-MMP2, anti-Cyclin B1 (Santa Cruz Biotechnology, Inc.), anti-cleaved caspase (Genetex), anti-heat shock factor 1 (HSF1), and anti-pHSF1 (Abcam, Inc.).

Techniques: Western Blot, Expressing, Staining, Flow Cytometry

Journal: Biomedicines

Article Title: Ponciri Fructus Immatarus Sensitizes the Apoptotic Effect of Hyperthermia Treatment in AGS Gastric Cancer Cells through ROS-Dependent HSP Suppression

doi: 10.3390/biomedicines11020405

Figure Lengend Snippet: Schematic diagram illustrating the present study. Bcl, B-cell lymphoma; ERK, extracellular signal-regulated kinase; HSF1, heat shock factor 1; HSP, heat shock protein; JNK, Jun N-terminal kinase; MAPK, mitogen-activated protein kinase; MMP, matrix metallopeptidase; NAC, N-acetylcysteine; PF, Ponciri Fructus Immaturus; ROS, reactive oxygen species; TNFR, tumor necrosis factor receptor; VEGF, vascular endothelial growth factor.

Article Snippet: After blocking, the matched primary antibodies (1:3000) were applied to the membranes, including the following primary antibodies at 4 °C overnight: anti-caspase-3, anti-survivin, anti-heat shock protein (HSP) 27, anti-HSP70, anti-HSP90, anti-caspase-8, anti-caspase-9, anti-p-extracellular signal-regulated kinase (ERK) (Thr202/Tyr204), anti-ERK, anti-p-Jun N-terminal kinase (JNK) (Thr183/Tyr185), anti-JNK, anti-p-p38 (Thr180/Tyr182), anti-p38 (Cell Signaling Technology), anti-β-actin, anti-Bcl-2, anti-Bcl-xL, anti-cyclin D1, anti-vascular endothelial growth factor (VEGF), anti-matrix metallopeptidase (MMP) 9, anti-MMP2, anti-Cyclin B1 (Santa Cruz Biotechnology, Inc.), anti-cleaved caspase (Genetex), anti-heat shock factor 1 (HSF1), and anti-pHSF1 (Abcam, Inc.).

Techniques:

Journal: PLoS ONE

Article Title: FOXO4-Knockdown Suppresses Oxidative Stress-Induced Apoptosis of Early Pro-Angiogenic Cells and Augments Their Neovascularization Capacities in Ischemic Limbs

doi: 10.1371/journal.pone.0092626

Figure Lengend Snippet: (A) Representative fluorescence microscopic images of EPCs incorporated in endothelial cells of the ischemic limbs of athymic nude rats 60 h after intramuscular injection of EPCs, NT-EPCs, or FOXO4 KD -EPCs. Rat specific CD31-positive endothelial cells were stained red. Human specific PCNA-positive cells, which were defined as human-derived EPCs, were stained green. Scale bar: 50 μm. (B) Pooled data of the number of PCNA-positive EPCs incorporated in endothelial cells of the rat ischemic limbs 60 h after the injection of EPCs, NT-EPCs, or FOXO4 KD -EPCs (*: p<0.01; **: p<0.005; n = 5, each). (C) A representative western blotting photo of expressions of human specific VEGF, b-FGF, SDF-1, and IGF-1 in the rat ischemic limb 60 h after intramuscular injection of EPCs, NT-EPCs, or FOXO4 KD -EPCs (n = 4, each).

Article Snippet: The membrane was blocked with a buffer (Blocking One, Nacalai Tesque, Inc.) and incubated for 24 h at 4°C with primary antibodies as follows: a rabbit anti-human β-actin antibody, a rabbit anti-human FOXO3a antibody, a rabbit anti-human FOXO4 antibody, a rabbit anti-human Bim antibody, a rabbit anti-human cleaved caspase-3 antibody, a rabbit anti-human vascular endothelial growth factor (VEGF) antibody, a rabbit anti-human basic-fibroblast growth factor (b-FGF) antibody (above antibodies were purchased from Cell Signaling Technology), a rabbit anti-human stromal cell-derived factor-1 (SDF-1) antibody, and a rabbit anti-human insulin-like growth factor-1 (IGF-1) antibody (above antibodies were purchased from abcam).

Techniques: Fluorescence, Injection, Staining, Derivative Assay, Western Blot

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Saposhnikoviae Radix Enhanced the Angiogenic and Anti-Inflammatory Effects of Huangqi Chifeng Tang in a Rat Model of Cerebral Infarction

doi: 10.1155/2021/4232708

Figure Lengend Snippet: Effects of HQCFT and HQCFT-SR on the protein expression levels of MMP-9, VEGF, and VEGFR2 in the brain tissue of rats subjected to MCAO. (a) MMP-9 expression in rat brain tissue, (b) VEGF expression in rat brain tissue, and (c) VEGFR2 expression in rat brain tissue. The sham group and model group were administered normal saline for 14 days. The HQCFT group and HQCFT-SR group were treated with HQCFT extract and HQCFT without SR for 14 days, respectively, and the dose was 1.6 g/kg and 1.31 g/kg daily, respectively. Data are expressed as the mean ± SD ( n = 3). ∗∗∗ p < 0.001 vs. the sham operation group, # p < 0.05, ## p < 0.01, and ### p < 0.001 vs. the model group.

Article Snippet: Subsequently, the membranes were blocked with 5% skim milk for 1 h at room temperature and incubated overnight at 4°C with the indicated primary antibodies against NLRP3 (1 : 500, Bios), caspase 1 (1 : 1000, Abcam), IL-1 β (1 : 500, Abcam), IL-6 (1 : 1000, Abcam), TNF- α (1 : 500, Gene Tex), MMP-9 (1 : 500, Bios), vascular endothelial growth factor (VEGF) (1 : 500, Bios), vascular endothelial growth factor receptor 2 (VEGFR2) (1 : 1000, CST), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1 : 2500, Abcam).

Techniques: Expressing

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Saposhnikoviae Radix Enhanced the Angiogenic and Anti-Inflammatory Effects of Huangqi Chifeng Tang in a Rat Model of Cerebral Infarction

doi: 10.1155/2021/4232708

Figure Lengend Snippet: Immunohistochemical staining of (a) MMP-9, (b) VEGF, and (c) VEGFR2 in rat brain tissue. (A) The sham group and (B) model group were administered normal saline for 14 days. (C) The HQCFT group and (D) HQCFT-SR group were treated with HQCFT extract and HQCFT without SR for 14 days, respectively, and the dose was 1.6 g/kg and 1.31 g/kg daily, respectively. The sections were magnified 200 times, and gray MMP-9-, VEGF-, and VEGFR2-positive cells were found in the rat brain tissue. The pictures shown are representative of three independent samples.

Article Snippet: Subsequently, the membranes were blocked with 5% skim milk for 1 h at room temperature and incubated overnight at 4°C with the indicated primary antibodies against NLRP3 (1 : 500, Bios), caspase 1 (1 : 1000, Abcam), IL-1 β (1 : 500, Abcam), IL-6 (1 : 1000, Abcam), TNF- α (1 : 500, Gene Tex), MMP-9 (1 : 500, Bios), vascular endothelial growth factor (VEGF) (1 : 500, Bios), vascular endothelial growth factor receptor 2 (VEGFR2) (1 : 1000, CST), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1 : 2500, Abcam).

Techniques: Immunohistochemical staining, Staining

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Saposhnikoviae Radix Enhanced the Angiogenic and Anti-Inflammatory Effects of Huangqi Chifeng Tang in a Rat Model of Cerebral Infarction

doi: 10.1155/2021/4232708

Figure Lengend Snippet: The levels of MMP-9, VEGF, and VEGFR2 in the rat brain tissue. (a) The optical densities of MMP-9-positive cells. (b) The optical densities of VEGF-positive cells. (c) The optical densities of VEGFR2-positive cells. Images were processed and analyzed with ImageJ software. Data are expressed as the mean ± SD ( n = 3). ∗∗∗ p < 0.001 vs. the sham group, ## p < 0.01 and ### p < 0.001 vs. the model group, && p < 0.01 and &&& p < 0.001 vs. the HQCFT group.

Article Snippet: Subsequently, the membranes were blocked with 5% skim milk for 1 h at room temperature and incubated overnight at 4°C with the indicated primary antibodies against NLRP3 (1 : 500, Bios), caspase 1 (1 : 1000, Abcam), IL-1 β (1 : 500, Abcam), IL-6 (1 : 1000, Abcam), TNF- α (1 : 500, Gene Tex), MMP-9 (1 : 500, Bios), vascular endothelial growth factor (VEGF) (1 : 500, Bios), vascular endothelial growth factor receptor 2 (VEGFR2) (1 : 1000, CST), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1 : 2500, Abcam).

Techniques: Software