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