Journal: Experimental and Therapeutic Medicine

Article Title: Expression and significance of NELIN and SM22α in varicose vein tissue

doi: 10.3892/etm.2015.2170

Figure Lengend Snippet: Primer sequences used for reverse transcription polymerase chain reaction analysis of NELIN.

Article Snippet: TransZol Up and two-step reverse transcription polymerase chain reaction (RT-PCR) kits were purchased from Beijing TransGen Biotech Co., Ltd. (Beijing, China). β-actin upstream and downstream primers were synthesized by Takara Bio, Inc. (Shiga, Japan).

Techniques: Reverse Transcription Polymerase Chain Reaction, Amplification

Journal: Experimental and Therapeutic Medicine

Article Title: Expression and significance of NELIN and SM22α in varicose vein tissue

doi: 10.3892/etm.2015.2170

Figure Lengend Snippet: Primer sequences used for reverse transcription polymerase chain reaction analysis of SM22α.

Article Snippet: TransZol Up and two-step reverse transcription polymerase chain reaction (RT-PCR) kits were purchased from Beijing TransGen Biotech Co., Ltd. (Beijing, China). β-actin upstream and downstream primers were synthesized by Takara Bio, Inc. (Shiga, Japan).

Techniques: Reverse Transcription Polymerase Chain Reaction, Amplification

Journal: Experimental and Therapeutic Medicine

Article Title: Expression and significance of NELIN and SM22α in varicose vein tissue

doi: 10.3892/etm.2015.2170

Figure Lengend Snippet: Reverse transcription polymerase chain reaction analysis revealed weak mRNA expression of NELIN in the vascular smooth muscle cells (VSMCs) of the experimental group, but strong expression in the control group. (A) Expected length of the PCR products was 611 bp (NELIN) and 312 bp (β-actin). Lanes: N, control group; X, DNA marker DL 2000; and P, experimental group. (B) Downregulation of NELIN mRNA expression in the VSMCs from the varicose vein tissue. Data are presented as the mean ± standard deviation.

Article Snippet: TransZol Up and two-step reverse transcription polymerase chain reaction (RT-PCR) kits were purchased from Beijing TransGen Biotech Co., Ltd. (Beijing, China). β-actin upstream and downstream primers were synthesized by Takara Bio, Inc. (Shiga, Japan).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Marker, Standard Deviation

Journal: Experimental and Therapeutic Medicine

Article Title: Expression and significance of NELIN and SM22α in varicose vein tissue

doi: 10.3892/etm.2015.2170

Figure Lengend Snippet: Reverse transcription polymerase chain reaction analysis revealed weak mRNA expression of SM22α in the vascular smooth muscle cells (VSMCs) of the experimental group, but strong expression in the control group. (A) Expected length of the PCR products was 232 bp (SM22α) and 472 bp (GADPH). Lanes: N, control group; X, DNA marker DL 2000; and P, experimental group. (B) Downregulation of SM22α mRNA expression in the VSMCs from the varicose vein tissue. Data are presented as the mean ± standard deviation.

Article Snippet: TransZol Up and two-step reverse transcription polymerase chain reaction (RT-PCR) kits were purchased from Beijing TransGen Biotech Co., Ltd. (Beijing, China). β-actin upstream and downstream primers were synthesized by Takara Bio, Inc. (Shiga, Japan).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Marker, Standard Deviation

Journal: Cellular and Molecular Immunology

Article Title: Chemerin aggravates DSS-induced colitis by suppressing M2 macrophage polarization

doi: 10.1038/cmi.2014.15

Figure Lengend Snippet: Increased production of pro-inflammatory cytokines in chemerin-treated mice following DSS exposure. ( a ) The concentrations of the pro-inflammatory cytokines IL-6, TNF-α and IFN-γ in the supernatant of colon cells after 24 h of culture were measured by ELISA. ( b ) The serum levels of the pro-inflammatory cytokines IL-6 and TNF-α were measured by ELISA. * P <0.05, ** P <0.01, *** P <0.001. The data are expressed as the mean±s.e.m. ( n =4–6 per group). Similar results were obtained in three independent experiments with 4–6 mice per group. DSS, dextran sulfate sodium; ELISA, enzyme-linked immunosorbent assay; IFN, interferon.

Article Snippet: The concentrations of chemerin were measured using an ELISA Kit for mouse chemerin (R&D Systems).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Cellular and Molecular Immunology

Article Title: Chemerin aggravates DSS-induced colitis by suppressing M2 macrophage polarization

doi: 10.1038/cmi.2014.15

Figure Lengend Snippet: Attenuated intestinal inflammation in neutralizing anti-chemerin (ChAb)-treated mice following DSS exposure. ( a ) The methods for DSS-induced colitis and ChAb administration. ( b ) Representative photomicrographs of colon sections stained with H&E were examined at ×40 magnification. ( c ) Histological scores were determined as described in the section on ‘Materials and methods'. ( d ) The percentages of neutrophils, DCs and macrophages in the colons were determined by flow cytometry as described above. ( e ) The concentrations of the pro-inflammatory cytokines IL-6, TNF-α and IFN-γ in the supernatant of colon cells after 24 h of culture were measured by ELISA. ( f ) The colonic expression of the M2 macrophage-associated genes Arg1, Ym1, FIZZ1 and IL-10 was examined by RT-PCR analysis. The data are expressed as the mean±s.e.m. ( n =5 per group). Similar results were obtained in three independent experiments with five mice per group. ns=not significant, * P <0.05, ** P <0.01, *** P <0.001. ChAb, chemerin antibody; DC, dendritic cell; DSS, dextran sulfate sodium; ELISA, enzyme-linked immunosorbent assay; IFN, interferon; UC, ulcerative colitis.

Article Snippet: The concentrations of chemerin were measured using an ELISA Kit for mouse chemerin (R&D Systems).

Techniques: Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Oncogene

Article Title: Increased metabolites of 5-lipoxygenase from hypoxic ovarian cancer cells promote tumor-associated macrophage infiltration.

doi: 10.1038/onc.2014.85

Figure Lengend Snippet: Figure 2. Key proteins and metabolites of the 5-LOX pathway are increased in hypoxic ovarian cancer cells. (a) SKOV-3 and OVCAR-3 ovarian cancer cells were treated with 1% O2 for 0, 4, 8, 12 and 24 h. Transcript levels of 5-LOX, FLAP and LTA4H were analyzed by quantitative reverse transcriptase-PCR. *Po0.05 versus 0 h. (b) SKOV-3 cells were treated with 1% O2 for 0, 6, 12, 24 and 48 h. The expression of HIF-1α, 5-LOX and FLAP were analyzed by western blotting. *Po0.05 versus Con. (c) SKOV-3 cells were treated with 1% O2 for 6, 12, 24, 48 and 72 h. CM was harvested and analyzed by ELISA. *Po0.05 versus normoxia. (d) SKOV-3 cells were transfected with siRNAs and treated with 1% O2 for 24 h. The expression of HIF-1α, 5-LOX and FLAP were analyzed by western blotting. *Po0.05 versus Con siRNA. #Po0.05 versus corresponding Con siRNA.

Article Snippet: For measurement of secreted HB-EGF and TNF-α, levels of HB-EGF and TNF-α present in the supernatants were measured using the human HB-EGF ELISA kit (Cusabio Biotech Co., Ltd) and human TNF-α ELISA Kit (R&D Systems) following the manufacture’s protocol.

Techniques: Reverse Transcription, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Transfection

Journal: Oncogene

Article Title: Increased metabolites of 5-lipoxygenase from hypoxic ovarian cancer cells promote tumor-associated macrophage infiltration.

doi: 10.1038/onc.2014.85

Figure Lengend Snippet: Figure 6. Metabolites of 5-LOX enhance release of TNF-α and HB-EGF through upregulation of MMP-7. TNF-α and HB-EGF in medium were detected by ELISA. *Po0.05 versus Con. #Po0.05 versus corresponding CM, CM+isotype-IgG or isotype-IgG. Macrophages were cultured with medium alone (Con), normoxia (N) or hypoxia (H) CM from SKOV-3 cells (a) or untreated and MK886-treated SKOV-3 cells (b). (c) Isotype-IgG or anti-MMP-7 were added in the CM. Macrophages were treated with 5-HETE (d) or LTB4 (e). (f) Macrophages were treated with 5-HETE or LTB4. Then isotype-IgG or anti-MMP-7 were added in the medium.

Article Snippet: For measurement of secreted HB-EGF and TNF-α, levels of HB-EGF and TNF-α present in the supernatants were measured using the human HB-EGF ELISA kit (Cusabio Biotech Co., Ltd) and human TNF-α ELISA Kit (R&D Systems) following the manufacture’s protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Frontiers in Toxicology

Article Title: Nicotine-free electronic vape fluid stimulates angiogenic processes in vitro through ARF6-mediated oxidative stress

doi: 10.3389/ftox.2025.1699112

Figure Lengend Snippet: Nicotine-free e-cigarette condensate (eVape) exposure significantly increases angiogenic processes in human endothelial cells. (a) Human umbilical vein endothelial cells (HUVECs) were exposed to eVape fluid containing 0, 5, and 10 mg/mL of nicotine for 24 h at concentrations ranging from 0% to 2%. Cell viability was measured using the MTT assay, quantified at 570 nm, and calculated as percentage viability normalised to vehicle. (b, c) Endothelial cells were exposed to 2% eVape containing 0 mg/mL of nicotine or vascular endothelial growth factor (VEGF; 100 ng/mL) or vehicle control (H 2 O) for 6 h. (b) Cell adhesion was measured using MTT, while (c) cell migration was assessed using scratch assay and calculated with MiToBo. (d–g) Neovascularisation was measured by culturing cells treated with 2% eVape containing 0 mg/mL of nicotine or VEGF (100 ng/mL) or vehicle control (H 2 O) on Matrigel TM -coated wells. Images were captured at ×20 magnification (scale bar: 100 µm; representative images are shown in (d) ). (e) Angiogenic potential, (f) tube formation, and (g) mesh size were quantified as number of joints, number of tubes, and pixel size, respectively, using Angiogenesis Analyser. The data are presented as mean ± standard error of the mean (SEM), n = 5–6. * p < 0.05 versus vehicle for eVape (0%); # p < 0.05 versus vehicle for VEGF (0 ng/mL).

Article Snippet: NAV2729 and Secin H3 were purchased from Tocris Bioscience (Abingdon, UK); Proteome Profiler Human angiogenesis array and enzyme-linked immunosorbent assay (ELISA) kits for human angiopoietin-2 (DANG20), endothelial growth factor (EGF; DEG00), endoglin (DNDG00), placental growth factor (PIGF; DPG00), prolactin (DY682), and vascular endothelial growth factor (VEGF; DVE00) were all obtained from R&D Systems (Abingdon, UK).

Techniques: MTT Assay, Control, Migration, Wound Healing Assay

Journal: Frontiers in Toxicology

Article Title: Nicotine-free electronic vape fluid stimulates angiogenic processes in vitro through ARF6-mediated oxidative stress

doi: 10.3389/ftox.2025.1699112

Figure Lengend Snippet: Nicotine-free eVape exposure causes significant upregulation of angiopoietin-2, endoglin, placental growth factor (PIGF), and VEGF as well as significant downregulation of endothelial growth factor (EGF) and prolactin in endothelial cells. HUVECs were exposed to 2% nicotine-free eVape fluid for 24 h. (a–c) Cell lysates were used for the Proteome Profiler Angiogenesis array membranes. Changes in the expression of all angiogenesis-related proteins in array (a) (ii) were quantified using the membranes (representative image in (a) (i) ). Specific analyses of the (b) upregulation and (c) downregulation of angiogenesis-related proteins on the membranes were quantified. (d) mRNA expression of angiopoietin-2, EGF, endoglin, PIGF, prolactin, and VEGF were measured by RT-PCR using specific primers. (e) Protein expression of (i) angiopoietin-2, (ii) EGF, (iii) endoglin, (iv) PIGF, (v) prolactin, and (vi) VEGF were measured using specific ELISA arrays. The data are presented as mean ± SEM, n = 5–6. * p < 0.05 versus vehicle for eVape (0%).

Article Snippet: NAV2729 and Secin H3 were purchased from Tocris Bioscience (Abingdon, UK); Proteome Profiler Human angiogenesis array and enzyme-linked immunosorbent assay (ELISA) kits for human angiopoietin-2 (DANG20), endothelial growth factor (EGF; DEG00), endoglin (DNDG00), placental growth factor (PIGF; DPG00), prolactin (DY682), and vascular endothelial growth factor (VEGF; DVE00) were all obtained from R&D Systems (Abingdon, UK).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Toxicology

Article Title: Nicotine-free electronic vape fluid stimulates angiogenic processes in vitro through ARF6-mediated oxidative stress

doi: 10.3389/ftox.2025.1699112

Figure Lengend Snippet: Activation of ARF6 by its guanine nucleotide exchange factor (ARNO) regulates angiogenic processes in endothelial cells induced by nicotine-free eVape. (a, b) HUVECs were exposed to 2% nicotine-free eVape fluid for 24 h, and the cell lysates were analysed by Western blotting for (a) ARF6 or (b) ARNO expression. The Western blotting membranes (representative blots are shown in (i) ) were quantified and normalised to the actin expression shown in (ii) . (c–f) HUVECs were treated with ARF6 inhibitor (NAV2729; 5 µM), ARNO inhibitor (Secin H3; 10 µM), or dimethyl sulfoxide (DMSO) as vehicle control at the same time as eVape exposure. (c) ROS accumulation was assessed by DCFDA incorporation (fluorescence at 485/535 nm) following 2 h of treatment. (d) Cell adhesion, (e) migration, and (f) angiogenic potential were measured following (d, e) 6 h and (f) 24 h of treatments. (g) Protein expression of (i) angiopoietin-2, (ii) EGF, (iii) endoglin, (iv) PIGF, (v) prolactin, and (vi) VEGF were measured using specific ELISA arrays following 24 h of treatment with 2% nicotine-free eVape fluid in the presence (closed circles) and absence (open circles) of Secin H3 (10 µM). The data are presented as mean ± SEM, n = 5–6. * p < 0.05 versus vehicle for eVape (0%); # p < 0.05 versus vehicle for NAV2729 and Secin H3 (DMSO).

Article Snippet: NAV2729 and Secin H3 were purchased from Tocris Bioscience (Abingdon, UK); Proteome Profiler Human angiogenesis array and enzyme-linked immunosorbent assay (ELISA) kits for human angiopoietin-2 (DANG20), endothelial growth factor (EGF; DEG00), endoglin (DNDG00), placental growth factor (PIGF; DPG00), prolactin (DY682), and vascular endothelial growth factor (VEGF; DVE00) were all obtained from R&D Systems (Abingdon, UK).

Techniques: Activation Assay, Western Blot, Expressing, Control, Fluorescence, Migration, Enzyme-linked Immunosorbent Assay

Journal: Scandinavian journal of immunology

Article Title: Chitinase induce the release of IL-8 in human airway epithelial cells, via Ca2+-dependent PKC and ERK pathways.

doi: 10.1111/j.1365-3083.2010.02404.x

Figure Lengend Snippet: Figure 2 The effect of mitogen-activated protein kinase and NF-kB on Streptomyces griseus chitinase-induced IL-8 production. (A) Cells treated with chitinase were harvested at the indicated time points and then lysed. The equal amounts of cell extracts were resolved on 10% acryl- amide gels and then subjected to Western blot analysis. (B) Cells were pre-treated with indicated concentrations of U0126, SB202190 and JNK inhibitor II for 1 h prior to stimulation with 10 lg ⁄ ml S. griseus chitinase. The supernatants were collected and evaluated for IL-8 pro- duction by ELISA. **P £ 0.001 versus chitinase alone. (C) Cells were pre-treated with different concentrations of caffeine acid phenethyl ester (CAPE) for 1 h prior to stimulation with 10 lg ⁄ ml S. griseus chitinase. The supernatants were collected and evaluated for IL-8 production. *P £ 0.05 versus chitinase alone. The data represent the mean ± SEM of three separate experiments.

Article Snippet: Cells were plated at 2 · 105 cells ⁄ ml in 12-well culture plates and stimulated with chitinase (2 and 10 lg ⁄ ml) for 2, 6, 12 or 24 h. The supernatants were harvested and measured to determine the level of IL-8 by human IL-8 ELISA kit (R&D system, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Scandinavian journal of immunology

Article Title: Chitinase induce the release of IL-8 in human airway epithelial cells, via Ca2+-dependent PKC and ERK pathways.

doi: 10.1111/j.1365-3083.2010.02404.x

Figure Lengend Snippet: Figure 1 The effect of chitinase on IL-8 production and mRNA expres- sion. (A) Cells were stimulated with different concentrations of Strepto- myces griseus chitinase (2 and 10 lg ⁄ ml) at 37 C. The supernatants were collected and assayed for IL-8 by ELISA. **P < 0.001 versus con- trol alone, *P < 0.01 versus control alone. The data represent the mean ± SEM of four separate experiments. (B) The RNAs from cells treated with S. griseus chitinase were harvested and used for RT-PCR.

Article Snippet: Cells were plated at 2 · 105 cells ⁄ ml in 12-well culture plates and stimulated with chitinase (2 and 10 lg ⁄ ml) for 2, 6, 12 or 24 h. The supernatants were harvested and measured to determine the level of IL-8 by human IL-8 ELISA kit (R&D system, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Reverse Transcription Polymerase Chain Reaction

Journal: Scandinavian journal of immunology

Article Title: Chitinase induce the release of IL-8 in human airway epithelial cells, via Ca2+-dependent PKC and ERK pathways.

doi: 10.1111/j.1365-3083.2010.02404.x

Figure Lengend Snippet: Figure 5 The effect of Streptomyces griseus chitinase on IL-6 production. Cells were stimulated with different concentrations of S. griseus chitinase (2 and 10 lg ⁄ ml). The supernatants were collected and assayed for IL-8 by ELISA. **P £ 0.01 versus control alone. The data represent the mean ± SEM of four separate experiments.

Article Snippet: Cells were plated at 2 · 105 cells ⁄ ml in 12-well culture plates and stimulated with chitinase (2 and 10 lg ⁄ ml) for 2, 6, 12 or 24 h. The supernatants were harvested and measured to determine the level of IL-8 by human IL-8 ELISA kit (R&D system, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Association of Serum HMGB2 Levels With In-Stent Restenosis

doi: 10.1161/atvbaha.116.308210

Figure Lengend Snippet: Figure 2. Hmgb2 deletion attenuates neointimal hyperplasia in injured femoral arteries of mice, whereas perivascular high-mobility group box (HMGB) 2 administration aggravates this pathology. A, Representative images of H&E staining of sham and wire-injured femoral artery sections from C57Bl/6 and Hmgb2–/– mice. Quantification of neointimal growth index (B) and intima-to-media (I/M) ratio (C) was performed in sham or injured arteries of C57Bl/6 and Hmgb2–/– mice. *P<0.05 vs injured arteries of C57Bl/6 mice; n=12. D, Immuno- fluorescent staining of α-smooth muscle actin (α-SMA) was performed in sections of injured arteries from C57Bl/6 and Hmgb2–/– mice. *P<0.05 vs injured arteries of C57Bl/6 mice; n=7. E, Expression of Col1a1, MMP2, and MMP9 in sham and injured arteries of C57Bl/6 and Hmgb2–/– mice. Data are quantified in Figure VI in the online-only Data Supplement. F, The mRNA levels of Col1a1, Col3a1, MMP2, and MMP9 in sham and injured arteries of C57Bl/6 and Hmgb2–/– mice as determined by reverse transcriptase polymerase chain reaction (RT- PCR). *P<0.05 vs injured arteries of C57Bl/6 mice; #P<0.05 vs sham arteries of C57Bl/6 or Hmgb2–/– mice; n=4. G, Representative images of H&E staining of sections of wire-injured arteries of PBS- or HMGB2-treated mice. H and I, Quantification of neointimal growth index (H) and intima-to-media (I/M) ratio (I) of wire-injured arteries of PBS- or HMGB2-treated mice. *P<0.05 vs PBS-treated group; n=10. J, Rep- resentative images of immunofluorescent staining for α-SMA performed in wire-injured arteries of PBS- or HMGB2-administrated mice (left). Staining was also quantified (right). *P<0.05 vs PBS-treated group; n=6 to 7. Masson’s trichrome staining of injury-induced femoral artery from C57Bl/6 and Hmgb2–/– mice (K) and PBS- or HMGB2-treated mice (L). Scale bar: 50 μm.

Article Snippet: The mice serum HMGB2 concentration was measured using a mouse HMGB2 ELISA kit (CSB-EL010560MO, CUSABIO Life Science) according to the manufacturer’s guidelines.

Techniques: Staining, Expressing, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Association of Serum HMGB2 Levels With In-Stent Restenosis

doi: 10.1161/atvbaha.116.308210

Figure Lengend Snippet: Figure 4. Reactive oxygen species (ROS) mediates high-mobility group box (HMGB) 2–promoted neointimal hyperplasia in mice. A, Rep- resentative images of dihydroethidium (DHE) staining in femoral arteries of C57Bl/6 or Hmgb2–/– mice with femoral artery wire injury or sham. B, Quantitative analysis of the DHE fluorescence intensity in arteries. #P<0.05 vs sham; *P<0.05 vs injured artery of C57Bl/6 mice; n=6. C, PF-127 gel containing PBS or HMGB2 was delivered around the femoral artery. Apocynin (APO) or vehicle was given in drinking water at a concentration of 1.5 mmol/L for 4 weeks. The sections of wire-injured arteries were stained with H&E, representative images are shown. Quantification of neointimal growth index (D) and intima-to-media (I/M) ratio (E). *P<0.05 vs PBS administration; #P<0.05 vs HMGB2 administration; n=10 to 11. F, Representative images of dihydroethidium (DHE) staining in wire-injured femoral arteries of C57Bl/6 mice with perivascular administration of HMGB2 or PBS, in the presence or absence of APO. G,: Quantitative analysis of the DHE fluores- cent intensity in arteries. *P<0.05 vs sham; #P<0.05 vs injured artery of C57Bl/6 mice; n=5. Scale bar: 50 μm.

Article Snippet: The mice serum HMGB2 concentration was measured using a mouse HMGB2 ELISA kit (CSB-EL010560MO, CUSABIO Life Science) according to the manufacturer’s guidelines.

Techniques: Staining, Fluorescence, Concentration Assay

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Association of Serum HMGB2 Levels With In-Stent Restenosis

doi: 10.1161/atvbaha.116.308210

Figure Lengend Snippet: Figure 5. P47phox phosphorylation–dependent reactive oxygen species (ROS) activation mediates high-mobility group box (HMGB) 2–induced effects in vivo and in vitro. A, Western blot analysis of phospho–p47phox levels in injured femoral arteries of C57Bl/6 and Hmgb2–/– mice and in sham arteries. B, Quantification of phospho–p47phox levels in A. #P<0.05 vs sham; *P<0.05 vs injured arteries of C57Bl/6; n=3. C, Immunofluorescent staining of phospho–p47phox (red) and α-smooth muscle actin (α-SMA; green) in sections of sham or wire injury fem- oral artery in C57Bl/6 and Hmgb2–/– mice. Representative images are shown. D, Quantification of phospho–p47phox levels in femoral arter- ies of C. #P<0.05 vs sham; *P<0.05 vs injured artery of C57Bl/6; n=5. E, Human aortic smooth muscle cells (hASMCs) were stimulated with HMGB2 for various times (0, 15, 30, and 60 minutes). Phospho–p47phox and total p47phox levels were determined using Western blot analy- sis, with quantification as given in Figure XIF in the online-only Data Supplement. F, H, I, and K, In hASMCs transfected with scramble-small interfering RNA (siRNA) or p47phox-siRNA for 24 h and then stimulated by HMGB2 or PBS in experiments, ROS, proliferation assay, Boyden chamber assay, and Wound-healing assay were performed, and respectively, quantified in G, J, and L. *P<0.05 vs scramble-siRNA with PBS stimulation; #P<0.05 vs scramble-siRNA with HMGB2 stimulation; n=3 to 6. Scale bar: 50 μm. DCF indicates 2′,7′-dichlorodihydrofluorescein.

Article Snippet: The mice serum HMGB2 concentration was measured using a mouse HMGB2 ELISA kit (CSB-EL010560MO, CUSABIO Life Science) according to the manufacturer’s guidelines.

Techniques: Phospho-proteomics, Activation Assay, In Vivo, In Vitro, Western Blot, Staining, Transfection, Small Interfering RNA, Proliferation Assay, Boyden Chamber Assay, Wound Healing Assay

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Association of Serum HMGB2 Levels With In-Stent Restenosis

doi: 10.1161/atvbaha.116.308210

Figure Lengend Snippet: Figure 6. High-mobility group box (HMGB) 2 promotes p47phox phosphorylation and migration and proliferation of vascular smooth muscle cells (VSMCs) via receptor of advanced glycation end products (RAGE) but not NOX4. A, Human aortic smooth muscle cells (HASMCs) were transfected with scramble-small interfering RNA (siRNA), Toll-like receptor (TLR) 2-siRNA, TLR4-siRNA, or RAGE-siRNA for 24 h, followed by stimulation with PBS or HMGB2. The levels of phospho–p47phox and p47phox were determined by Western blot analysis. Quantification can be found in Figure XIII in the online-only Data Supplement. B and C, VSMCs derived from Rage–/– and Tlr4–/– mice were treated with PBS or HMGB2. The levels of phospho–p47phox and p47phox were determined by Western blot analysis. Quanti- fication can be found in Figure XIII in the online-only Data Supplement. D, F, and H, VSMCs derived from C57Bl/6 and Rage–/– mice were treated with PBS or HMGB2 in experiments. Reactive oxygen species (ROS), Boyden chamber, and proliferation assay were performed, and respectively, quantified in E and G. *P<0.05 vs C57Bl/6 mouse-derived VSMCs; #P<0.05 vs C57Bl/6 mouse-derived VSMCs with HMGB2 stimulation; n=3 to 5. I, Immunofluorescence confocal microscopy showed increased expression of HMGB2 and RAGE and colo- calization of these 2 proteins in neointima after vascular injury. n=4. Scale bar: 50 μm.

Article Snippet: The mice serum HMGB2 concentration was measured using a mouse HMGB2 ELISA kit (CSB-EL010560MO, CUSABIO Life Science) according to the manufacturer’s guidelines.

Techniques: Phospho-proteomics, Migration, Transfection, Small Interfering RNA, Western Blot, Derivative Assay, Proliferation Assay, Immunofluorescence, Confocal Microscopy, Expressing

Journal: Antioxidants (Basel, Switzerland)

Article Title: Skin Improvement with Antioxidant Effect of Yuja ( Citrus junos ) Peel Fractions: Wrinkles, Moisturizing, and Whitening.

doi: 10.3390/antiox12010051

Figure Lengend Snippet: Figure 1. Anti-wrinkle effects of YJP-EA, Hex, and BuOH on UVB-irradiated human keratinocyte HaCaT cells. (A) HaCaT cells were treated with YJP-EA, Hex, and BuOH for 24 h. An MTT assay was performed to evaluate cell viability. (B) HaCaT cells were irradiated with UVB (30 mJ/cm2) and then treated with YJP-EA, Hex, and BuOH for 24 h. Whole cell lysates were analyzed by Western blot analysis. (C) HaCaT cells were treated by YJP-EA, Hex, and BuOH for 24 h after UVB (30 mJ/cm2) irradiation. The RNA level was evaluated using reverse transcription PCR. (D) The Pro-Collagen of HaCaT cells was measured using an ELISA kit, following the manufacturer’s instructions, with 450 nm. All experiments were performed individually in triplicate. *** p < 0.001 vs. non-treated (NT) cells, ** p < 0.01 vs. non-treated (NT) cells, and * p < 0.05 vs. non-treated (NT) cells.

Article Snippet: The Type I Pro-Collagen amount was determined using a Human Pro-Collagen I α1 ELISA kit from R&D Systems (Minneapolis, MN, USA).

Techniques: Irradiation, MTT Assay, Western Blot, Reverse Transcription, Enzyme-linked Immunosorbent Assay

Journal: Antioxidants (Basel, Switzerland)

Article Title: Skin Improvement with Antioxidant Effect of Yuja ( Citrus junos ) Peel Fractions: Wrinkles, Moisturizing, and Whitening.

doi: 10.3390/antiox12010051

Figure Lengend Snippet: Figure 2. Anti-wrinkle effects of YJP-EA, Hex, and BuOH on UVB-irradiated human dermal fibroblast cells. (A) HDF cells were treated with YJP-EA, Hex, and BuOH for 24 h. An MTT assay was performed to evaluate cell viability. (B) HDF cells were irradiated with UVB (100 mJ/cm2) and then treated with YJP-EA, Hex, and BuOH for 24 h. Whole cell lysates were analyzed by Western blot analysis. (C) HDF cells were treated by YJP-EA, Hex, and BuOH for 24 h after UVB (100 mJ/cm2) irradiation. The RNA level was evaluated using reverse transcription PCR. (D) The Pro-Collagen of HDF cells was measured using an ELISA kit, following the manufacturer’s instructions, with 450 nm. All experiments were performed individually in triplicate. *** p < 0.001 vs. non-treated (NT) cells, ** p < 0.01 vs. non-treated (NT) cells, and * p < 0.05 vs. non-treated (NT) cells.

Article Snippet: The Type I Pro-Collagen amount was determined using a Human Pro-Collagen I α1 ELISA kit from R&D Systems (Minneapolis, MN, USA).

Techniques: Irradiation, MTT Assay, Western Blot, Reverse Transcription, Enzyme-linked Immunosorbent Assay