Journal: Biomedical Reports

Article Title: Knockdown of ALOX15 alleviates acute coronary syndrome via the FGFR2/PI3K/AKT signaling pathway

doi: 10.3892/br.2025.2092

Figure Lengend Snippet: Effects of ALOX15 on the FGFR2/PI3K/AKT signaling pathway in HCAECs. (A) Co-immunoprecipitation assays of ALOX15 and FGFR2 in HCAECs. (B) The mRNA expression of ALOX15 and FGFR2 in HCAECs after transfection with OE-ALOX15/OE-NC or siRNA ALOX15/siRNA NC was detected by reverse transcription-quantitative PCR. *** P<0.001. (C) The protein levels of ALOX15, FGFR2, PI3K, p-PI3K, AKT and p-AKT in HCAECs after transfection with OE-ALOX15/OE-NC or siRNA ALOX15/siRNA NC were determined by western blotting. *** P<0.001. ALOX15, 12/15-lipoxygenase; FGFR2, fibroblast growth factor receptor 2; HCAECs, human primary coronary artery endothelial cells; OE, overexpression; NC, negative control; siRNA, small interfering RNA; p-, phosphorylated; NS, not significant.

Article Snippet: TRIzol reagent (cat. no. 10606ES60), Hifair ® II 1st Strand cDNA Synthesis SuperMix (cat. no. 11123ES60) and Hieff ® qPCR SYBR Green Master Mix (cat. no. 11202ES08) were procured from Shanghai Yeasen Biotechnology Co., Ltd. Primary antibodies AKT (cat. no. 60203-2-Ig), phoshorylated (p)-AKT (cat. no. 66444-1-Ig), GAPDH (cat. no. 60004-1-Ig) and HRP-conjugated secondary antibodies (cat. no. SA00001-2) were obtained from Proteintech Group, Inc. Primary antibodies p-PI3K (cat. no. 4228T) and FGFR2 (cat. no. 23328) were purchased from Cell Signaling Technology, Inc., and PI3K antibody (cat. no. ab191606) was sourced from Abcam.

Techniques: Immunoprecipitation, Expressing, Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Over Expression, Negative Control, Small Interfering RNA

Journal: Biomedical Reports

Article Title: Knockdown of ALOX15 alleviates acute coronary syndrome via the FGFR2/PI3K/AKT signaling pathway

doi: 10.3892/br.2025.2092

Figure Lengend Snippet: Silencing of ALOX15 mitigates acute coronary syndrome progression in vitro through the FGFR2/PI3K/AKT signaling pathway. (A) The expression of FGFR2 in HCAECs after transfection with OE-FGFR2 or OE-NC was detected by RT-qPCR. (B) The mRNA expression of ALOX15 and FGFR2 in HCAECs following different treatments was detected by RT-qPCR. (C) The protein levels of FGFR2, PI3K, p-PI3K, AKT and p-AKT in HCAECs following various treatments were determined by western blotting. (D, E) The migration of HCAECs following different treatments was assessed by scratch wound healing assays. Scale bar, 100 µm. (F) The viability of HCAECs following various treatments was measured by Cell Counting Kit-8 assays. * P<0.05, ** P<0.01 and *** P<0.001. ALOX15, 12/15-lipoxygenase; FGFR2, fibroblast growth factor receptor 2; HCAECs, human primary coronary artery endothelial cells; OE, overexpression; NC, negative control; RT-qPCR, reverse transcription-quantitative PCR; siRNA, small interfering RNA; p-, phosphorylated; NS, not significant.

Article Snippet: TRIzol reagent (cat. no. 10606ES60), Hifair ® II 1st Strand cDNA Synthesis SuperMix (cat. no. 11123ES60) and Hieff ® qPCR SYBR Green Master Mix (cat. no. 11202ES08) were procured from Shanghai Yeasen Biotechnology Co., Ltd. Primary antibodies AKT (cat. no. 60203-2-Ig), phoshorylated (p)-AKT (cat. no. 66444-1-Ig), GAPDH (cat. no. 60004-1-Ig) and HRP-conjugated secondary antibodies (cat. no. SA00001-2) were obtained from Proteintech Group, Inc. Primary antibodies p-PI3K (cat. no. 4228T) and FGFR2 (cat. no. 23328) were purchased from Cell Signaling Technology, Inc., and PI3K antibody (cat. no. ab191606) was sourced from Abcam.

Techniques: In Vitro, Expressing, Transfection, Quantitative RT-PCR, Western Blot, Migration, Cell Counting, Over Expression, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Small Interfering RNA

Journal: Biomedical Reports

Article Title: Knockdown of ALOX15 alleviates acute coronary syndrome via the FGFR2/PI3K/AKT signaling pathway

doi: 10.3892/br.2025.2092

Figure Lengend Snippet: ALOX15 silencing suppresses the activation of the FGFR2/PI3K/AKT signaling pathway. The protein levels of ALOX15, FGFR2, PI3K, p-PI3K, AKT and p-AKT in rats with ACS after injection of shRNA ALOX15 or shRNA NC were determined by western blotting. *** P<0.001 vs. sham; ### P<0.001 vs. ACS model + shRNA NC. ALOX15, 12/15-lipoxygenase; FGFR2, fibroblast growth factor receptor 2; p-, phosphorylated; ACS, acute coronary syndrome; shRNA, short hairpin RNA; NC, negative control; NS, no significance.

Article Snippet: TRIzol reagent (cat. no. 10606ES60), Hifair ® II 1st Strand cDNA Synthesis SuperMix (cat. no. 11123ES60) and Hieff ® qPCR SYBR Green Master Mix (cat. no. 11202ES08) were procured from Shanghai Yeasen Biotechnology Co., Ltd. Primary antibodies AKT (cat. no. 60203-2-Ig), phoshorylated (p)-AKT (cat. no. 66444-1-Ig), GAPDH (cat. no. 60004-1-Ig) and HRP-conjugated secondary antibodies (cat. no. SA00001-2) were obtained from Proteintech Group, Inc. Primary antibodies p-PI3K (cat. no. 4228T) and FGFR2 (cat. no. 23328) were purchased from Cell Signaling Technology, Inc., and PI3K antibody (cat. no. ab191606) was sourced from Abcam.

Techniques: Activation Assay, Injection, shRNA, Western Blot, Negative Control

Journal: iScience

Article Title: 3D-printed scaffold loaded with baicalin exosomes promotes bone defect repair via mediating PRRX2 to alleviate inflammation

doi: 10.1016/j.isci.2025.113565

Figure Lengend Snippet: BA-BMSC-exos inhibited inflammation via PRRX2/IL-6/AKT pathway (A) The heatmap of RNA-seq data illustrates the differentially expressed genes. (B) KEGG pathway enrichment. (C) The mRNA level of PRRX2 in HUVECs. (D) Immunofluorescence analysis of PRRX2 in HUVECs (scale bar: 100 μm). (E) Western blot analysis of p -AKT in HUVECs. (F) The binding of Prrx2 to the IL-6 gene promoter was studied by using the ChIP method, and the promoter of IL-6 was amplified by qPCR. (G) Luciferase activity of wild or mutant IL-6 was tested by luciferase reporter assay. (H) Hypothetical scheme for the mechanisms of PRRX2 inhibiting inflammation. Data are presented as mean ± standard deviation (SD), n = 3, p -values are calculated using one-way ANOVA, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: p -AKT Antibody , Proteintech , Cat # 66444-1-Ig; RRID: AB_2782958.

Techniques: RNA Sequencing, Immunofluorescence, Western Blot, Binding Assay, Amplification, Luciferase, Activity Assay, Mutagenesis, Reporter Assay, Standard Deviation