Journal: International journal of molecular medicine

Article Title: FGF21 attenuates hypoxia‑induced dysfunction and apoptosis in HPAECs through alleviating endoplasmic reticulum stress.

doi: 10.3892/ijmm.2018.3705

Figure Lengend Snippet: Figure 4. FGF21 alleviates the hypoxia‑induced ERS by modulating the expression of ERS‑related proteins in HPAECs. HPAECs were treated as indicated and protein expression levels were examined by western blotting. Representative blots and quantification are shown for (A) BiP, (B) p‑PERK and PERK, (C) the ratio of p‑PERK/PERK, (D) CHOP, (E) Bcl‑2. (F) Relative expression of the ERS‑dependent apoptotic protein caspase‑4 in HPAECs was analysed by western blotting. GAPDH was used as an internal control. All experiments were performed in triplicate, and data are expressed as the mean ± standard deviation (n=3). **P<0.01 vs. N group; ##P<0.01 vs. the H group; &&P<0.01 vs. the H+T group. FGF21, fibroblast growth factor 21; ERS, endoplasmic reticulum stress; HPAECs, human pulmonary arterial endothelial cells; BiP, binding immunoglobulin protein; p‑, phosphorylated; PERK, protein kinase R‑like endoplasmic reticulum kinase; CHOP, transcrip- tion factor C/EBP homologous protein; Bcl‑2, B cell lymphoma-2; N, normoxia; H, hypoxia; F, FGF21; S, salubrinal; T, tunicamycin.

Article Snippet: Rabbit antibodies against phosphorylated (p-) PERK were purchased from Biorbyt (cambridge, UK).

Techniques: Expressing, Western Blot, Control, Standard Deviation, Binding Assay

Journal: International journal of molecular medicine

Article Title: FGF21 attenuates hypoxia‑induced dysfunction and apoptosis in HPAECs through alleviating endoplasmic reticulum stress.

doi: 10.3892/ijmm.2018.3705

Figure Lengend Snippet: Figure 6. Effect of FGF21 on NO and ET‑1 secretion in HPAECs. (A) HPAECs were treated as indicated and at the end of treatment, the cell culture medium was collected and assayed by ELISA for the levels of secreted NO and (B) the levels of secreted ET‑1. Data are expressed as the mean ± standard deviation (n=3). **P<0.01 vs. the N group; #P<0.05 and ##P<0.01 vs. the H group; &&P<0.01 vs. the H+T group. (C) Schematic of the proposed mechanism by which FGF21 attenuates hypoxia‑induced apoptosis and dysfunction by alleviating ERS in HPAECs. FGF21, fibroblast growth factor 21; NO, nitric oxide; ET‑1, endothelin‑1; HPAECs, human pulmonary arterial endothelial cells; ERS, endoplasmic reticulum stress; N, normoxia; H, hypoxia; F, FGF21; S, salubrinal; T, tunicamycin; BiP, binding immunoglobulin protein; p‑, phosphorylated; PERK, protein kinase R‑like endoplasmic reticulum kinase; CHOP, transcription factor C/EBP homologous protein; Bcl‑2, B cell lymphoma-2.

Article Snippet: Rabbit antibodies against phosphorylated (p-) PERK were purchased from Biorbyt (cambridge, UK).

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Standard Deviation, Binding Assay

Journal: Frontiers in Immunology

Article Title: Suppression of Endoplasmic Reticulum Stress by 4-PBA Protects Against Hyperoxia-Induced Acute Lung Injury via Up-Regulating Claudin-4 Expression

doi: 10.3389/fimmu.2021.674316

Figure Lengend Snippet: 4-PBA alleviates hyperoxia-induced ER stress-related proteins in lung tissues. (A) Western blot analysis of GRP78, p-PERK, p-IRE1, ATF6, CHOP, ATF4, and p-elF-2 expression in lung tissues. Relative expressions of GRP78 (B) , p-PERK (C) , p-IRE1 (D) , ATF6 (E) , CHOP (F) , ATF4 (G) , and p-elF-2 (H) in lung tissues were shown. Data are expressed as mean ± SD (3 mice per group). **p < 0.01, ***p < 0.001, compared with the room air group; + p < 0.05, ++ p < 0.01 compared with the hyperoxia group.

Article Snippet: The membranes were blocked for 1 hours at room temperature with 5% nonfat milk and then incubated overnight at 4°C with the following antibodies: claudin-3 (Invitrogen, Carlsbad, CA, USA), claudin-4 (OriGene Technologies, Inc., Rockville, MD, USA), claudin-18 (Invitrogen), GRP78 (Abcam, Cambridge, MA, USA), phospho PERK (Taiclone, Taipei, Taiwan), phospho IRE1 (Abcam), ATF6 (Bioss Antibodies, Woburn, MA, USA), ATF4 (Bioss), GADD153 (CHOP, Santa Cruz Biotechnology, Dallas, TX, USA), B-cell lymphoma (Bcl)-2 (Santa Cruz Biotechnology), NF-κB p65, phospho-NF-κB p65, inhibitor of NF-κB (IκB)-α, cleaved caspase-3 and proliferating cell nuclear antigen (PCNA, Cell Signaling Technology), and β-actin (Sigma-Aldrich).

Techniques: Western Blot, Expressing

Journal: Frontiers in Immunology

Article Title: Suppression of Endoplasmic Reticulum Stress by 4-PBA Protects Against Hyperoxia-Induced Acute Lung Injury via Up-Regulating Claudin-4 Expression

doi: 10.3389/fimmu.2021.674316

Figure Lengend Snippet: Schematic shows that hyperoxia triggers endoplasmic reticulum (ER) stress responses that lead to inflammation and apoptosis. The inflammation and apoptosis impair tight junction function and lung vascular permeability that induce acute lung injury. 4-PBA administration improves these phenomena. ATF4, activating transcription factor 4; ATF6, activating transcription factor 6; IRE1, inositol-requiring enzyme 1; PERK, protein kinase-like ER kinase. p-elF-2α, eukaryotic translation initiation factor 2α.

Article Snippet: The membranes were blocked for 1 hours at room temperature with 5% nonfat milk and then incubated overnight at 4°C with the following antibodies: claudin-3 (Invitrogen, Carlsbad, CA, USA), claudin-4 (OriGene Technologies, Inc., Rockville, MD, USA), claudin-18 (Invitrogen), GRP78 (Abcam, Cambridge, MA, USA), phospho PERK (Taiclone, Taipei, Taiwan), phospho IRE1 (Abcam), ATF6 (Bioss Antibodies, Woburn, MA, USA), ATF4 (Bioss), GADD153 (CHOP, Santa Cruz Biotechnology, Dallas, TX, USA), B-cell lymphoma (Bcl)-2 (Santa Cruz Biotechnology), NF-κB p65, phospho-NF-κB p65, inhibitor of NF-κB (IκB)-α, cleaved caspase-3 and proliferating cell nuclear antigen (PCNA, Cell Signaling Technology), and β-actin (Sigma-Aldrich).

Techniques: Permeability