Journal: Frontiers in Pharmacology

Article Title: ANGPTL4 Regulates Psoriasis via Modulating Hyperproliferation and Inflammation of Keratinocytes

doi: 10.3389/fphar.2022.850967

Figure Lengend Snippet: Role of ANGPTL4 in the IMQ-induced psoriasiform dermatitis in mice. (A) Schematic illustration of the animal experiment protocol for the CON, IMQ, IMQ + A4 group ( n = 5). (B) Phenotypical presentation of mouse back skin and light microscopy examination of skin sections stained with H&E after 7 days of CON, IMQ, IMQ + A4 groups. Scale bars, 50 μm. (C) Daily assessment of epidermal erythema, scales, and thickness of the shaved backs. PASI score was calculated to assess the severity of psoriasis by adding the scores of three criteria (range from 0 to 12). (D) The skin thickness was measured on day 8. Significant differences are indicated. (E,F) Representative immunohistochemical stainings and quantification of ANGPTL4 in skin sections. Scale bars, 50 μm. A4, ANGPTL4; CON group = 62.5 mg/day Vaseline cream +25 ug/kg/day vehicle i. d. for 7 days, IMQ group = 62.5 mg/day 5% IMQ cream +25ug/kg/day vehicle i. d., IMQ + A4 group = 62.5 mg 5% IMQ cream+ 25 ug/kg/day recombinant ANGPTL4 i. d. Data were shown as mean ± standard deviation (SD); * p < 0.05, ** p < 0.01, *** p < 0.001 vs. CON group.

Article Snippet: After blocking in 5% bovine albumin (BSA) for 1 h, membranes were incubated with primary antibodies at against STAT3 (1:1,000, A1192, ABclonal), p-STAT3-Y705 (1:1,000, AP0705, ABclonal), ERK1/ERK2 (1:1,000, A16686, ABclonal), p-ERK1-T202/Y204 + ERK2-T185/Y187 (1:1,000, AP0472, ABclonal), PCNA (1:1,000, A0264, ABclonal), Cyclin D1 (1:1,000, 2922S, Cell Signaling Technology) Cleaved IL-1β (1:1,000, Cell Signaling Technology), IL-17A (1:1,000, A0688, ABclonal), ANGPTL4 (1:1,000, CSB-PA005044, Cusabio), ANGPTL4 (1:500, AF3485, R&D Systems), GAPDH (1:5,000, CSB-MA000071M2m, Cusabio) at 4°C overnight.

Techniques: Light Microscopy, Staining, Immunohistochemical staining, Recombinant, Standard Deviation

Journal: Frontiers in Pharmacology

Article Title: ANGPTL4 Regulates Psoriasis via Modulating Hyperproliferation and Inflammation of Keratinocytes

doi: 10.3389/fphar.2022.850967

Figure Lengend Snippet: Identification of modules associated with psoriasis using the WGCNA. (A) Gene dendrogram of the top 25% most variant genes clustered based on a dissimilarity measure. In total, 12 modules were identified. (B) Module eigengene dendrogram and eigengene adjacency heatmap summarize the modules yielded in the hierarchical clustering analysis. (C) Heatmap of the module-trait correlation. (D) Volcano plot of genes detected in psoriasis. Red means up-regulated DEGs; blue means down-regulated DEGs; grey means no difference. (E) Venn diagrams of the intersection between DEGs and the hub genes in the turquoise module. (F) Box plot showing the differences in ANGPTL4 gene expression between lesional and paired non-lesional skin in the same patient. LS, psoriasis lesions skin; NL, non-lesional skin; DEG, differentially expressed gene.

Article Snippet: After blocking in 5% bovine albumin (BSA) for 1 h, membranes were incubated with primary antibodies at against STAT3 (1:1,000, A1192, ABclonal), p-STAT3-Y705 (1:1,000, AP0705, ABclonal), ERK1/ERK2 (1:1,000, A16686, ABclonal), p-ERK1-T202/Y204 + ERK2-T185/Y187 (1:1,000, AP0472, ABclonal), PCNA (1:1,000, A0264, ABclonal), Cyclin D1 (1:1,000, 2922S, Cell Signaling Technology) Cleaved IL-1β (1:1,000, Cell Signaling Technology), IL-17A (1:1,000, A0688, ABclonal), ANGPTL4 (1:1,000, CSB-PA005044, Cusabio), ANGPTL4 (1:500, AF3485, R&D Systems), GAPDH (1:5,000, CSB-MA000071M2m, Cusabio) at 4°C overnight.

Techniques: Variant Assay, Expressing

Journal: Frontiers in Pharmacology

Article Title: ANGPTL4 Regulates Psoriasis via Modulating Hyperproliferation and Inflammation of Keratinocytes

doi: 10.3389/fphar.2022.850967

Figure Lengend Snippet: ANGPTL4 expression is elevated in IMQ-induced psoriasiform dermatitis in mice. IMQ or control cream (vaseline) was applied daily to the dorsal skin for female BALB/c mice. (A) Phenotypical presentation of mouse back skin and light microscopy examination of skin sections stained with H&E after 7 days of control cream (upper panel) or IMQ treatment (lower panel). Scale bars, 50 μm. (B) The skin thickness was measured on day 8. Significant differences are indicated ( n = 5). (C) Daily assessment of epidermal erythema, scales, and thickeness of the shaved backs. PASI score was calculated to assess the severity of psoriasis by adding the scores of three criteria (range from 0 to 12). (D,E) Representative immunoblotting and analysis of ANGPTL4 from skin samples of control cream or IMQ treated mice on day 8 ( n = 6). (F) Relative mRNA expression of ANGPTL4 in IMQ treatment mice and control cream on day 8 ( n = 5). (G,H) Representative immunohistochemical stainings and quantification of ANGPTL4 in skin sections ( n = 5). Scale bars, 50 μm. (I,J) Representative images and quantification of ANGPTL4 fluorescence in skin sections. The nuclei were detected by DAPI (blue). Scale bar, 50 μm. All data were shown as mean ± standard deviation (SD). IMQ, imiquimod; H&E, haematoxylin and eosin; IOD, the integral optical density. * p < 0.05, ** p < 0.01 vs. CON group.

Article Snippet: After blocking in 5% bovine albumin (BSA) for 1 h, membranes were incubated with primary antibodies at against STAT3 (1:1,000, A1192, ABclonal), p-STAT3-Y705 (1:1,000, AP0705, ABclonal), ERK1/ERK2 (1:1,000, A16686, ABclonal), p-ERK1-T202/Y204 + ERK2-T185/Y187 (1:1,000, AP0472, ABclonal), PCNA (1:1,000, A0264, ABclonal), Cyclin D1 (1:1,000, 2922S, Cell Signaling Technology) Cleaved IL-1β (1:1,000, Cell Signaling Technology), IL-17A (1:1,000, A0688, ABclonal), ANGPTL4 (1:1,000, CSB-PA005044, Cusabio), ANGPTL4 (1:500, AF3485, R&D Systems), GAPDH (1:5,000, CSB-MA000071M2m, Cusabio) at 4°C overnight.

Techniques: Expressing, Light Microscopy, Staining, Western Blot, Immunohistochemical staining, Fluorescence, Standard Deviation

Journal: Frontiers in Pharmacology

Article Title: ANGPTL4 Regulates Psoriasis via Modulating Hyperproliferation and Inflammation of Keratinocytes

doi: 10.3389/fphar.2022.850967

Figure Lengend Snippet: ANGPTL4 expression is elevated in psoriasis patients. (A) Phenotypical presentation of psoriasis and normal skin stained with H&E. Scale bars, 250 μm. (B) The skin thickness was measured. Significant differences are indicated ( n = 5). (C,D) Representative immunohistochemical stainings and quantification of ANGPTL4 in skin sections from psoriasis and non-psoriasis patients ( n = 5). Scale bars, 250 μm. (E,F) Representative images and quantification of ANGPTL4 fluorescence in skin sections. The nuclei were detected by DAPI (blue). Scale bar, 100 μm. All data were shown as mean ± standard deviation (SD). H&E, haematoxylin and eosin; IOD, the integral optical density. ** p < 0.01 vs. CON group.

Article Snippet: After blocking in 5% bovine albumin (BSA) for 1 h, membranes were incubated with primary antibodies at against STAT3 (1:1,000, A1192, ABclonal), p-STAT3-Y705 (1:1,000, AP0705, ABclonal), ERK1/ERK2 (1:1,000, A16686, ABclonal), p-ERK1-T202/Y204 + ERK2-T185/Y187 (1:1,000, AP0472, ABclonal), PCNA (1:1,000, A0264, ABclonal), Cyclin D1 (1:1,000, 2922S, Cell Signaling Technology) Cleaved IL-1β (1:1,000, Cell Signaling Technology), IL-17A (1:1,000, A0688, ABclonal), ANGPTL4 (1:1,000, CSB-PA005044, Cusabio), ANGPTL4 (1:500, AF3485, R&D Systems), GAPDH (1:5,000, CSB-MA000071M2m, Cusabio) at 4°C overnight.

Techniques: Expressing, Staining, Immunohistochemical staining, Fluorescence, Standard Deviation

Journal: Frontiers in Pharmacology

Article Title: ANGPTL4 Regulates Psoriasis via Modulating Hyperproliferation and Inflammation of Keratinocytes

doi: 10.3389/fphar.2022.850967

Figure Lengend Snippet: Effect of ANGPTL4 on Human Keratinocyte Cells (A) Representative Western blots showing PCNA, Cyclin D1, Cleaved IL-1β and IL-17A in HaCaT cells after treatment with ANGPTL4 (500 ng/ml). (B) WB analysis of PCNA, Cyclin D1, Cleaved IL-1β, IL-17A expression, respectively. (C) CCK-8 assay results ( n = 10). (D–G) Relative mRNA expression of IL-1β, IL-17A, IL-6 and TNF-α normalized to a GAPDH internal control ( n = 5). Values are expressed relative to control groups. (H–K) The secretion of IL-1β, IL-17, IL-6 and TNF-α were determined by ELISA ( n = 3). Results shown are representative data of three independent experiments. All data were shown as mean ± standard deviation (SD). ** p < 0.01, *** p < 0.001 vs. CON group.

Article Snippet: After blocking in 5% bovine albumin (BSA) for 1 h, membranes were incubated with primary antibodies at against STAT3 (1:1,000, A1192, ABclonal), p-STAT3-Y705 (1:1,000, AP0705, ABclonal), ERK1/ERK2 (1:1,000, A16686, ABclonal), p-ERK1-T202/Y204 + ERK2-T185/Y187 (1:1,000, AP0472, ABclonal), PCNA (1:1,000, A0264, ABclonal), Cyclin D1 (1:1,000, 2922S, Cell Signaling Technology) Cleaved IL-1β (1:1,000, Cell Signaling Technology), IL-17A (1:1,000, A0688, ABclonal), ANGPTL4 (1:1,000, CSB-PA005044, Cusabio), ANGPTL4 (1:500, AF3485, R&D Systems), GAPDH (1:5,000, CSB-MA000071M2m, Cusabio) at 4°C overnight.

Techniques: Western Blot, Expressing, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Frontiers in Pharmacology

Article Title: ANGPTL4 Regulates Psoriasis via Modulating Hyperproliferation and Inflammation of Keratinocytes

doi: 10.3389/fphar.2022.850967

Figure Lengend Snippet: Effect of ANGPTL4 Knockdown on Human Keratinocyte Cells (A,B) Representative Western blots and quantification of ANGPTL4, PCNA, Cyclin D1, Cleaved IL-1β and IL-17A in cultured HaCaT cells under the treatment with si-RNA targeting ANGPTL4 (si-ANG-01 and si-ANG-02) or negative control siRNA (si-NC). GAPDH was used as loading control ( n = 3). (C) CCK-8 assay results ( n = 9). (D) Relative mRNA expression of IL-1β, IL-17A, IL-6 and TNF-α normalized to a GAPDH internal control ( n = 5). Results shown are representative data of three independent experiments. All data were shown as mean ± standard deviation (SD). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. CON group.

Article Snippet: After blocking in 5% bovine albumin (BSA) for 1 h, membranes were incubated with primary antibodies at against STAT3 (1:1,000, A1192, ABclonal), p-STAT3-Y705 (1:1,000, AP0705, ABclonal), ERK1/ERK2 (1:1,000, A16686, ABclonal), p-ERK1-T202/Y204 + ERK2-T185/Y187 (1:1,000, AP0472, ABclonal), PCNA (1:1,000, A0264, ABclonal), Cyclin D1 (1:1,000, 2922S, Cell Signaling Technology) Cleaved IL-1β (1:1,000, Cell Signaling Technology), IL-17A (1:1,000, A0688, ABclonal), ANGPTL4 (1:1,000, CSB-PA005044, Cusabio), ANGPTL4 (1:500, AF3485, R&D Systems), GAPDH (1:5,000, CSB-MA000071M2m, Cusabio) at 4°C overnight.

Techniques: Western Blot, Cell Culture, Negative Control, CCK-8 Assay, Expressing, Standard Deviation

Journal: Frontiers in Pharmacology

Article Title: ANGPTL4 Regulates Psoriasis via Modulating Hyperproliferation and Inflammation of Keratinocytes

doi: 10.3389/fphar.2022.850967

Figure Lengend Snippet: Modulation of STAT3 and ERK1/2 pathways by ANGPTL4 in HaCaT cells. (A) Representative Western blots showing p-ERK1/2, ERK1/2, p-STAT3, STAT3 levels in HaCaT cells after treatment with ANGPTL4 (500 ng/ml). (B) Representative Western blots showing PCNA, Cyclin D1, Cleaved IL-1β and IL-17A in HaCaT cells after treatment with ANGPTL4 and MEK inhibitor (PD0325901). (C) Representative Western blots showing p-ERK1/2, ERK1/2, p-STAT3 and STAT3 levels in HaCaT cells under treatment with si-RNA targeting ANGPTL4 (si-ANG-01 and si-ANG-02) or negative control siRNA (si-NC). GAPDH was used as loading control ( n = 3). (D) Quantitative Western blot analysis. Results shown are representative data of three independent experiments. All data were shown as mean ± standard deviation (SD). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. CON group.

Article Snippet: After blocking in 5% bovine albumin (BSA) for 1 h, membranes were incubated with primary antibodies at against STAT3 (1:1,000, A1192, ABclonal), p-STAT3-Y705 (1:1,000, AP0705, ABclonal), ERK1/ERK2 (1:1,000, A16686, ABclonal), p-ERK1-T202/Y204 + ERK2-T185/Y187 (1:1,000, AP0472, ABclonal), PCNA (1:1,000, A0264, ABclonal), Cyclin D1 (1:1,000, 2922S, Cell Signaling Technology) Cleaved IL-1β (1:1,000, Cell Signaling Technology), IL-17A (1:1,000, A0688, ABclonal), ANGPTL4 (1:1,000, CSB-PA005044, Cusabio), ANGPTL4 (1:500, AF3485, R&D Systems), GAPDH (1:5,000, CSB-MA000071M2m, Cusabio) at 4°C overnight.

Techniques: Western Blot, Negative Control, Standard Deviation

Journal: Frontiers in Pharmacology

Article Title: ANGPTL4 Regulates Psoriasis via Modulating Hyperproliferation and Inflammation of Keratinocytes

doi: 10.3389/fphar.2022.850967

Figure Lengend Snippet: Role of ANGPTL4 in the IMQ-induced psoriasiform dermatitis in mice. (A,B) Representative Western blots and quantification showing phospho-ERK1/2, ERK1/2, phospho-STAT3, STAT3, Cleaved IL-1β and IL-17A levels in the CON, IMQ, IMQ + A4 group ( n = 3). (C) Relative mRNA expression of IL-1β, IL-17A, IL-6 and TNF-α in the CON, IMQ, IMQ + A4 group ( n = 5). Data were shown as mean ± standard deviation (SD); ** p < 0.01, *** p < 0.001 vs. CON group.

Article Snippet: After blocking in 5% bovine albumin (BSA) for 1 h, membranes were incubated with primary antibodies at against STAT3 (1:1,000, A1192, ABclonal), p-STAT3-Y705 (1:1,000, AP0705, ABclonal), ERK1/ERK2 (1:1,000, A16686, ABclonal), p-ERK1-T202/Y204 + ERK2-T185/Y187 (1:1,000, AP0472, ABclonal), PCNA (1:1,000, A0264, ABclonal), Cyclin D1 (1:1,000, 2922S, Cell Signaling Technology) Cleaved IL-1β (1:1,000, Cell Signaling Technology), IL-17A (1:1,000, A0688, ABclonal), ANGPTL4 (1:1,000, CSB-PA005044, Cusabio), ANGPTL4 (1:500, AF3485, R&D Systems), GAPDH (1:5,000, CSB-MA000071M2m, Cusabio) at 4°C overnight.

Techniques: Western Blot, Expressing, Standard Deviation

Journal: Oncology reports

Article Title: Expression of angiopoietin-like 4 (ANGPTL4) in human colorectal cancer: ANGPTL4 promotes venous invasion and distant metastasis.

doi: 10.3892/or.2011.1176

Figure Lengend Snippet: Figure 1. Immunohistochemical staining for ANGPTL4 in human tissues. (A) Faintly expression in the surface lining cells of normal colon mucosa (arrow- heads), (B) strongly in cytoplasm of moderately-differentiated adenocarcinoma, and (C) poorly-differentiated adenocarcinoma (non-solid type) of human colorectal cancer (magnification, x100).

Article Snippet: Deparaffinized sections were preincubated with normal rabbit serum to prevent non-specific binding, and then incubated overnight at 4 ̊C with an optimal dilution (0.1 μg/ml) of a primary polyclonal goat antibody against human Angptl4 (r&D systems, Inc., minneapolis, mA, usA). the slides were sequentially incubated with a biotinylated rabbit antigoat immunoglobulin antibody, and the reaction products were viewed using diaminobenzidine (DAb; Dako ltd.) and counterstained with hematoxylin. primary antibody preabsorbed with excess recombinant Angptl4 peptides (r&D systems, Inc.) was used as negative control.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Oncology reports

Article Title: Expression of angiopoietin-like 4 (ANGPTL4) in human colorectal cancer: ANGPTL4 promotes venous invasion and distant metastasis.

doi: 10.3892/or.2011.1176

Figure Lengend Snippet: Figure 2. Overall survival based on the expression of ANGPTL4 in human colorectal cancer (60 cases). ANGPTL4 expression was not associated with the overall survival in univariate survival analysis. (p=0.455 in log-rank test).

Article Snippet: Deparaffinized sections were preincubated with normal rabbit serum to prevent non-specific binding, and then incubated overnight at 4 ̊C with an optimal dilution (0.1 μg/ml) of a primary polyclonal goat antibody against human Angptl4 (r&D systems, Inc., minneapolis, mA, usA). the slides were sequentially incubated with a biotinylated rabbit antigoat immunoglobulin antibody, and the reaction products were viewed using diaminobenzidine (DAb; Dako ltd.) and counterstained with hematoxylin. primary antibody preabsorbed with excess recombinant Angptl4 peptides (r&D systems, Inc.) was used as negative control.

Techniques: Expressing

Journal: Oncology reports

Article Title: Expression of angiopoietin-like 4 (ANGPTL4) in human colorectal cancer: ANGPTL4 promotes venous invasion and distant metastasis.

doi: 10.3892/or.2011.1176

Figure Lengend Snippet: Figure 3. RT-PCR and Western blot analysis for ANGPTL4 in human colo rectal cancer: tissues and cultured cell lines. (A) RT-PCR. M, 100 bp ladder marker (Invitrogen, Inc.). (B) Western blot analysis. Human colorectal cancer tissues (1-6) and human colorectal cancer cell lines (7-12). [1, Cancer tissue; cases 1 and 2, normal gastric tissue; cases 1 and 3, cancer tissue; cases 2 and 4, normal gastric tissue; cases 2, 5, cancer tissue; cases 3 and 6, normal gastric tissue; cases-3 and 7, Caco-2; 8, Colo201; 9, Colo320DM; 10, DLD-1; 11, WiDr,; 12, LS123].

Article Snippet: Deparaffinized sections were preincubated with normal rabbit serum to prevent non-specific binding, and then incubated overnight at 4 ̊C with an optimal dilution (0.1 μg/ml) of a primary polyclonal goat antibody against human Angptl4 (r&D systems, Inc., minneapolis, mA, usA). the slides were sequentially incubated with a biotinylated rabbit antigoat immunoglobulin antibody, and the reaction products were viewed using diaminobenzidine (DAb; Dako ltd.) and counterstained with hematoxylin. primary antibody preabsorbed with excess recombinant Angptl4 peptides (r&D systems, Inc.) was used as negative control.

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Cell Culture, Marker

Journal: Journal of Lipid Research

Article Title: Characterization of ANGPTL4 function in macrophages and adipocytes using Angptl4 -knockout and Angptl4 -hypomorphic mice [S]

doi: 10.1194/jlr.M094128

Figure Lengend Snippet: ANGPTL4 deficiency increases respiration in BMDMs but does not influence LPL abundance. A: Real-time changes in OCR in BMDMs from Angptl4+/+ and Angptl4−/− mice in the presence or absence of recombinant ANGPTL4, treated with 0.5 mM intralipid for 6 h, and assessed in unbuffered medium during sequential injections with oligomycin, FCCP, and antimycin A + rotenone. B: Basal (left) and maximal (right) OCR in BMDMs from Angptl4+/+ and Angptl4−/− mice in the presence or absence of recombinant ANGPTL4 (0.5 μg/ml) incubated with 0.5 mM intralipid for 6 h. Measurements were done in quintuplicates in unbuffered medium. Data are mean ± SD; ***P < 0.001. C: Immunoblot of LPL in Angptl4+/+ and Angptl4−/− BMDMs treated for 6 h with or without 0.5 mM intralipid in the presence or absence of endonuclease-H. D: Immunoblot of LPL from culture medium of Angptl4+/+ and Angptl4−/− BMDMs treated with or without heparin. E: Immunoblot of LPL in Angptl4+/+ and Angptl4−/− BMDMs treated with or without ER to Golgi transport inhibitors monensin (Mon; 10 μM for 3 h) and Brefeldin A (BFA; 5 μg/ml for 4 h), proteasomal degradation inhibitor MG132 (40 μM for 4 h), and lysosomal degradation inhibitors, e64d (20 μM for 24 h) and leupeptin (Leup; 5 μM for 16 h). HSP90 or Coomassie staining serve as loading control.

Article Snippet: Membranes were probed with a goat anti-mouse LPL antibody ( 41 ), a rabbit anti-mouse HSP90 antibody (#4874S; Cell Signaling), a rat anti-mouse ANGPTL4 antibody (Kairos 142-2; Adipogen), and a rabbit anti-mouse ANGPTL4 antibody (#742, home made) ( 42 ) at 1:5,000 (LPL), 1:2,000 (HSP90), or 1:1,000 (ANGPTL4) dilutions.

Techniques: Recombinant, Incubation, Western Blot, Staining, Control

Journal: Molecular medicine reports

Article Title: Hydrogen peroxide promotes the expression of angiopoietin like 4 in RAW264.7 macrophages via MAPK pathways.

doi: 10.3892/mmr.2017.7365

Figure Lengend Snippet: Figure 2. H2O2 activates ERK1/2, p38 MAPK and JNK protein phosphorylation expression in RAW264.7 macrophages. Cells were stimulated with 0 (control), 0.25 or 0.5 mM H2O2 for 24 h, then protein expression in cell lysates was measured by western blotting. Cell lysates were probed for (A) p‑ and total ERK1/2, (B) p‑ and total p38 MAPK and (C) p‑ and total JNK, with α‑tubulin as loading control. *P<0.05, **P<0.01 and ***P<0.001 vs. control. ERK1/2, MAPK1; MAPK, mitogen‑activated protein kinase; JNK, MAPK8; p‑, phosphorylated.

Article Snippet: Primary antibody against ANGPTL4 (cat. no. 40-9800) was obtained from Invitrogen; Thermo Fisher Scientific, Inc. Primary antibodies against phosphorylated (p-)p38 MAPK (cat. no. 4511), p38 MAPK (cat. no. 8690), p-ERK1/2 (cat. no. 4370), ERK1/2 (cat. no. 4695), p-JNK (cat. no. 4668) and JNK (cat. no. 9252) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA), as were U0126 (cat. no. 9903) and SB203580 (cat. no. 5633).

Techniques: Phospho-proteomics, Expressing, Control, Western Blot

Journal: Molecular medicine reports

Article Title: Hydrogen peroxide promotes the expression of angiopoietin like 4 in RAW264.7 macrophages via MAPK pathways.

doi: 10.3892/mmr.2017.7365

Figure Lengend Snippet: Figure 3. Effects of MAPK inhibitors on H2O2‑induced ANGPTL4 release in RAW264.7 macrophage cells. Cells were pretreated with specific inhibitors for ERK1/2 (20 mM U0126), p38 MAPK (40 mM SB203580) or JNK (10 mM SP600125), followed by stimulation with 0, 0.25 or 0.5 mM H2O2 for 24 h. Protein expression levels were then determined by western blotting, with α‑tubulin as loading control. (A) ANGPTL4, p‑ERK1/2 and total ERK1/2. (B) ANGPTL4, p‑p38 MAPK and total p38 MAPK. (C) ANGPTL4, p‑JNK and total JNK. *P<0.05, **P<0.01 and ***P<0.001, with comparisons indicated by brackets. MAPK, mitogen‑activated protein kinase; ANGPTL4, angiopoietin like 4; ERK1/2, MAPK1; JNK, MAPK8; p‑, phosphorylated; Control, 0 mM H2O2 and no inhibitor; ns, not significant.

Article Snippet: Primary antibody against ANGPTL4 (cat. no. 40-9800) was obtained from Invitrogen; Thermo Fisher Scientific, Inc. Primary antibodies against phosphorylated (p-)p38 MAPK (cat. no. 4511), p38 MAPK (cat. no. 8690), p-ERK1/2 (cat. no. 4370), ERK1/2 (cat. no. 4695), p-JNK (cat. no. 4668) and JNK (cat. no. 9252) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA), as were U0126 (cat. no. 9903) and SB203580 (cat. no. 5633).

Techniques: Expressing, Western Blot, Control