Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Alpha-tocopherol inhibits ferroptosis and promotes neural function recovery in rats with spinal cord injury via downregulating Alox15.

doi: 10.1016/j.biopha.2024.116734

Figure Lengend Snippet: Fig. 3. Alpha-tocopherol alleviates ferroptosis after SCI. (A-B) Prussian Blue Iron Staining (Enhance With DAB) and quantitation. The positive stained tissue was observed in SCI group. The positive cells (iron overloaded cells) were colored by yellow-brown, the negative cells (normal cells) were colored by purple or blue. (C-F) WB and quantitation of Alox15, Ptgs2 and 4Hne in vivo. (G-J) IF and quantitation of Alox15 and Ptgs2. NeuN was colored by red, Alox15 and Ptgs2 were colored by green. n = 5 for each group. Data are represented as the means ± SD. ***p < 0.001.

Article Snippet: Slices were incubated overnight at 4°C with the primary antibodies against Alox15 (rabbit, 1:200, Affinity, Cat# DF13494, RRID: AB_2846513), Ptgs2 (rabbit, 1:200, Affinity, Cat# AF7003, RRID: AB_2835311), Gapdh (mouse, 1:200, Proteintech, Wuhan, China, Cat# 60004–1-Ig, RRID: AB_2107436) and neuronal nuclei (NeuN, mouse, 1:200, Proteintech, Cat# 66836–1-Ig, RRID: AB_2882179).

Techniques: Staining, Quantitation Assay, In Vivo

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Alpha-tocopherol inhibits ferroptosis and promotes neural function recovery in rats with spinal cord injury via downregulating Alox15.

doi: 10.1016/j.biopha.2024.116734

Figure Lengend Snippet: Fig. 6. Alpha-tocopherol downregulates expression of Alox15 and Ptgs2 in PC12 cells. (A-B) QPCR for mRNA of Alox15 and Ptgs2. (C-F) WB and quantitation of Alox15, Ptgs2 and 4Hne in vitro. (G-J) IF and quantitation of Alox15 and Ptgs2. Alox15 and Ptgs2 were colored by green. n = 3 for each group. Data are represented as the means ± SD. **p < 0.01, ***p < 0.001.

Article Snippet: Slices were incubated overnight at 4°C with the primary antibodies against Alox15 (rabbit, 1:200, Affinity, Cat# DF13494, RRID: AB_2846513), Ptgs2 (rabbit, 1:200, Affinity, Cat# AF7003, RRID: AB_2835311), Gapdh (mouse, 1:200, Proteintech, Wuhan, China, Cat# 60004–1-Ig, RRID: AB_2107436) and neuronal nuclei (NeuN, mouse, 1:200, Proteintech, Cat# 66836–1-Ig, RRID: AB_2882179).

Techniques: Expressing, Quantitation Assay, In Vitro

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Alpha-tocopherol inhibits ferroptosis and promotes neural function recovery in rats with spinal cord injury via downregulating Alox15.

doi: 10.1016/j.biopha.2024.116734

Figure Lengend Snippet: Fig. 7. Site-directed mutagenesis on Alox15 reverses protective effects of alpha-tocopherol. (A-B) WB validation of Alox15 knockout. (C) Erastin IC50 for Alox15-KO cells after transfection with different plasmids. (D) 48 hours after transfection, cells were treated with 5 μM erastin for 12 hours, with or without treatment of alpha- tocopherol or liproxstatin-1. Cell viabilities were detected by cck-8 assays. (E) 48 hours after transfection, cells were treated with 5 μM erastin for 12 hours, with or without treatment of alpha-tocopherol or liproxstatin-1. Alox15, Ptgs2 and 4Hne expressions were detected by WB. (F-H) Quantitation of WB. (I) Lipid oxidation detected by C11-BODIPY (581/591) probe. Green indicates oxidized lipids and red indicates non-oxidized lipids. (J) Quantitation of C11. n = 3 for each group. Data are represented as the means ± SD. ns: no significance, ***p < 0.001.

Article Snippet: Slices were incubated overnight at 4°C with the primary antibodies against Alox15 (rabbit, 1:200, Affinity, Cat# DF13494, RRID: AB_2846513), Ptgs2 (rabbit, 1:200, Affinity, Cat# AF7003, RRID: AB_2835311), Gapdh (mouse, 1:200, Proteintech, Wuhan, China, Cat# 60004–1-Ig, RRID: AB_2107436) and neuronal nuclei (NeuN, mouse, 1:200, Proteintech, Cat# 66836–1-Ig, RRID: AB_2882179).

Techniques: Mutagenesis, Biomarker Discovery, Knock-Out, Transfection, CCK-8 Assay, Quantitation Assay

Journal: Diabetology & Metabolic Syndrome

Article Title: Didymin protects pancreatic beta cells by enhancing mitochondrial function in high-fat diet-induced impaired glucose tolerance

doi: 10.1186/s13098-023-01244-1

Figure Lengend Snippet: Antibodies

Article Snippet: MT-CO2 , Rabbit , Cell Signaling Technology , #31,219.

Techniques:

Journal: Diabetology & Metabolic Syndrome

Article Title: Didymin protects pancreatic beta cells by enhancing mitochondrial function in high-fat diet-induced impaired glucose tolerance

doi: 10.1186/s13098-023-01244-1

Figure Lengend Snippet: Didymin improves mitochondrial function in PBCs. ( A ) MitoTracker Green staining for mitochondrial content in INS-1 cells. Scale bar = 30 μm. (n = 4) ( B ) Western blot analysis of NRF1, TFAM, NDUFB8, SDHB and MTCO2 in INS-1 cells (n = 3). ( C ) Mitochondrial oxygen consumption ratio (OCR) of INS-1 cells (n = 4). ( D ) ROS concentration in INS-1 cells (n = 4). ( E ) Analysis of mitochondrial structure of PBCs in mice by electron microscopy. Scale bar = 0.6 μm. ( F ) Immunofluorescence staining of COX-IV (green) and insulin (red) in pancreas. Scale bar = 20 μm. Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01, **** P < 0.0001 PA vs. PA + Didymin. # P < 0.05, ## P < 0.01, ### P < 0.001, #### P < 0.0001 control vs. PA.

Article Snippet: MT-CO2 , Rabbit , Cell Signaling Technology , #31,219.

Techniques: Staining, Western Blot, Concentration Assay, Electron Microscopy, Immunofluorescence, Control