Journal: Scientific Reports

Article Title: MiR-204-5p mediates PERK inhibition to suppress growth and induce apoptosis in ovarian cancer through the eIF2α/ATF-4/CHOP pathway

doi: 10.1038/s41598-025-95883-1

Figure Lengend Snippet: MiR-204-5p inhibited the phosphorylation of eIF2α, which is a downstream target of PERK. SKOV3 and OVCAR3 cells were transfected with either miR-204-5p mimics or negative control (NC) mimics for 24 h. Following this, the cells were treated with Tm at a concentration of 2 µg/ml. After 48 h, the phosphorylation levels of eIF2α were assessed using western blot analysis with specific antibodies in both OVCAR3 ( A ) and SKOV3 ( B ) cells. The results are presented as mean ± SD ( n = 3). ** p < 0.01; ## p < 0.01 compared to the untreated control group.

Article Snippet: The antibodies used in the western blot assays were as follows: PERK (C33E10; Cell Signaling; 1:500), B-cell lymphoma-2 (Bcl-2; sc-23960; Santa Cruz; 1:750), Bcl-2-associated X protein (Bax; sc-7480; Santa Cruz; 1:750), eIF2α (D7D3; Cell Signaling; 1:500), phosphorylated eIF2α (Ser 51; D9D8; Cell Signaling; 1:500), and CHOP (L63F7; Cell Signaling; 1:750), β- actin (sc-69879; Santa Cruz; 1:1000).

Techniques: Phospho-proteomics, Transfection, Negative Control, Concentration Assay, Western Blot, Control

Journal: Scientific Reports

Article Title: Hepatocyte-specific GDF15 overexpression improves high-fat diet-induced obesity and hepatic steatosis in mice via hepatic FGF21 induction

doi: 10.1038/s41598-024-75107-8

Figure Lengend Snippet: Effect of GDF15 overexpression on FGF21 expression in the liver for 8 weeks. ( A ) Hepatic Fgf21 mRNA levels were determined by quantitative real-time PCR ( n = 5–6/group). ( B ) Serum FGF21 levels were determined at the end of treatment ( n = 5–6/group). ( C , D ) Hepatic mRNA levels of Cnot6l ( C ) and unspliced and spliced Xbp1 ( Xbp1u and XBP1s ) ( D ) were determined by quantitative real-time PCR. Splicing efficiency (percentage of Xbp1s mRNA to Xbp1u mRNA) are shown. ( E , F ) PPARα ( E ) and p-eIF2a, total (t) eIF2α, XBP1s, and XBP1u levels ( F ) were evaluated by immunoblotting analysis of livers from 3 mice/group. Immunoblots were scanned, and band intensities were quantified using densitometric analysis. Each ratio was also expressed as fold change compared with controls. * P < 0.05, ** P < 0.01.

Article Snippet: Whole-liver protein lysates were separated using SDS-PAGE and transferred to polyvinylidene fluoride membranes, which were probed with primary antibodies to eIF2α, p-eIF2α, XBP1s, Erk1/2, or phosphorylated Erk1/2 (p-Erk1/2) from Cell Signaling Technology (Beverly, MA), PPARα (GTX101098) from GeneTex (Irvine, CA), unspliced XBP1 (clone F-4; sc-8015 F-4) from Santa Cruz (Dallas, TX, USA), GDF15 (bs-3818R) from Bioss (Boston, MA), and actin from Sigma Aldrich.

Techniques: Over Expression, Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Scientific Reports

Article Title: Hepatocyte-specific GDF15 overexpression improves high-fat diet-induced obesity and hepatic steatosis in mice via hepatic FGF21 induction

doi: 10.1038/s41598-024-75107-8

Figure Lengend Snippet: Effect of GDF15 overexpression on liver FGF21 expression after 2 weeks of induction. ( A ) Hepatic Gdf15 mRNA levels were determined by quantitative real-time PCR ( n = 5–6/group). Serum GDF15 levels were determined at the end of 2-week treatment ( n = 3–5/group). ( B ) Hepatic mRNA levels of Fgf21 and serum FGF21 levels were determined at the end of 2-week treatment ( n = 3–5/group). ( C , D , E ) PPARα ( C ), p-eIF2a/total (t) eIF2α ( D ), and p-Erk1/2/total (t) Erk12 ( E ) levels were evaluated by immunoblotting analysis of livers from 3 mice/group. Immunoblots were scanned, and band intensities were quantified using densitometric analysis. Each ratio was also expressed as a fold change compared with that of control. * P < 0.05, ** P < 0.01.

Article Snippet: Whole-liver protein lysates were separated using SDS-PAGE and transferred to polyvinylidene fluoride membranes, which were probed with primary antibodies to eIF2α, p-eIF2α, XBP1s, Erk1/2, or phosphorylated Erk1/2 (p-Erk1/2) from Cell Signaling Technology (Beverly, MA), PPARα (GTX101098) from GeneTex (Irvine, CA), unspliced XBP1 (clone F-4; sc-8015 F-4) from Santa Cruz (Dallas, TX, USA), GDF15 (bs-3818R) from Bioss (Boston, MA), and actin from Sigma Aldrich.

Techniques: Over Expression, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: Scientific Reports

Article Title: Hepatocyte-specific GDF15 overexpression improves high-fat diet-induced obesity and hepatic steatosis in mice via hepatic FGF21 induction

doi: 10.1038/s41598-024-75107-8

Figure Lengend Snippet: Effect of GDF15 overexpression in AML12 cells. ( A ) Microscopic images of lipid droplets and reactive oxygen species (ROS) based on a fluorescent dye in AML12 cells transfected with pLIVE-control or pLIVE-GDF15 before treatment with free fatty acids (FFAs) (scale bar = 50 μm [top] and 100 μm [bottom]). Representative photographs are shown. ( B , C ) Gdf15 , Fgf21 , and Cnot6l. ( D ) Cnot6l , Gdf15 , and Fgf21 . ( E ) Fgf21 and Cnot6l. ( F ) Xbp1s and Xbp1u mRNA levels were determined by quantitative real-time PCR ( n = 3). Splicing efficiencies are shown. ( G ) PPARα, p-eIF2a, total (t) eIF2α, XBP1s, XBP1u, and GDF15 levels were evaluated by immunoblotting. Representative blots from three independently repeated experiments are shown. Immunoblots were scanned, and band intensities were quantified using densitometric analysis. * P < 0.05, ** P < 0.01.

Article Snippet: Whole-liver protein lysates were separated using SDS-PAGE and transferred to polyvinylidene fluoride membranes, which were probed with primary antibodies to eIF2α, p-eIF2α, XBP1s, Erk1/2, or phosphorylated Erk1/2 (p-Erk1/2) from Cell Signaling Technology (Beverly, MA), PPARα (GTX101098) from GeneTex (Irvine, CA), unspliced XBP1 (clone F-4; sc-8015 F-4) from Santa Cruz (Dallas, TX, USA), GDF15 (bs-3818R) from Bioss (Boston, MA), and actin from Sigma Aldrich.

Techniques: Over Expression, Transfection, Control, Real-time Polymerase Chain Reaction, Western Blot