Journal: Neuroscience letters

Article Title: Mangiferin alleviates 6-OHDA-induced Parkinson's disease by inhibiting AKR1C3 to activate Wnt signaling pathway.

doi: 10.1016/j.neulet.2023.137608

Figure Lengend Snippet: Fig. 2. The inhibition of AKR1C3 activation of the Wnt signaling pathway in PC12 Cells by mangiferin. (A) Assessment of AKR1C3 expression levels using RT-qPCR. (B) Molecular docking simulations to investigate the binding affinity and interaction sites between mangiferin and AKR1C3 protein. (C) Measurement of gene expression levels of GSK-3β, Wnt1, and β-catenin using RT-qPCR. (D) Western blot analysis to examine the protein expression levels of p-GSK-3β, GSK-3β, Wnt1, β-catenin, and AKR1C3. *P<0.05, **P<0.01, ***P<0.001 vs the control group, #P<0.05, ##P<0.01, ###P<0.001, 6-OHDA+MG (50 μM) group vs 6-OHDA group, n=3. 6-OHDA: 6- hydroxydopamine, 100 μmol/L; MG: mangiferin, 50 μmol/L, one-way ANOVA.

Article Snippet: Primary antibodies are listed as follow: TH (1:5000, 25859-1-AP, proteintech), α-synuclein (1:1000, ab212184, Abcam), phosphate- GSK-3β (1:1000, AF2016, Affinity), GSK-3β (1:2000, ab93926, Abcam), AKR1C3 (1:2000, 11194-1-AP, proteintech), wnt1 (1:2000, 27935-1-AP, proteintech), β-catenin (1:10000, 51067-2-AP, proteintech), caspase-3 (1:2000, ab184787, Abcam), GAPDH (1:10000, ab8245, Abcam).

Techniques: Inhibition, Activation Assay, Expressing, Quantitative RT-PCR, Binding Assay, Gene Expression, Western Blot, Control

Journal: Neuroscience letters

Article Title: Mangiferin alleviates 6-OHDA-induced Parkinson's disease by inhibiting AKR1C3 to activate Wnt signaling pathway.

doi: 10.1016/j.neulet.2023.137608

Figure Lengend Snippet: Fig. 3. Weakening of the Protective Effect of mangiferin on PD by AKR1C3 Overexpression in PC12 Cells. (A) The influence of AKR1C3 overexpression on mitochondrial membrane potential using JC-1 staining. (B) The impact of AKR1C3 overexpression on intracellular reactive oxygen species (ROS) levels. (C) The effects of AKR1C3 overexpression on TH protein and α-synuclein protein. *P<0.05, **P<0.01, ***P<0.001 vs the control group, #P<0.05, ##P<0.01, ###P<0.001, 6- OHDA+MG (50 μM) group vs 6-OHDA group, n=3. PD: Parkinson’s disease; ROS, reactive oxygen species; TH: tyrosine hydroxylase; 6-OHDA: 6-hydroxydopamine, 100μmol/L; oeNC: over-expression negtive control; MG: mangiferin, 50μmol/L, one- way ANOVA.

Article Snippet: Primary antibodies are listed as follow: TH (1:5000, 25859-1-AP, proteintech), α-synuclein (1:1000, ab212184, Abcam), phosphate- GSK-3β (1:1000, AF2016, Affinity), GSK-3β (1:2000, ab93926, Abcam), AKR1C3 (1:2000, 11194-1-AP, proteintech), wnt1 (1:2000, 27935-1-AP, proteintech), β-catenin (1:10000, 51067-2-AP, proteintech), caspase-3 (1:2000, ab184787, Abcam), GAPDH (1:10000, ab8245, Abcam).

Techniques: Over Expression, Membrane, Staining, Control

Journal: Neuroscience letters

Article Title: Mangiferin alleviates 6-OHDA-induced Parkinson's disease by inhibiting AKR1C3 to activate Wnt signaling pathway.

doi: 10.1016/j.neulet.2023.137608

Figure Lengend Snippet: Fig. 4. Suppression of AKR1C3-mediated activation of the Wnt signaling by mangiferin in PC12 Cells. (A) Measurement of gene expression levels of GSK-3β, Wnt1, and mangiferin-catenin in AKR1C3-overexpressing cells using RT-qPCR. (B) Analysis of the protein expression levels of p-GSK-3mangiferin ,GSK-3β,Wnt1, and β- catenin through Western blotting in AKR1C3-overexpressing cells. *P<0.05, **P<0.01, ***P<0.001 vs the control group, #P<0.05, ##P<0.01, ###P<0.001, 6-OHDA+MG (50 μM) group vs 6-OHDA group, n=3. PD: Parkinson’s disease; TH: tyrosine hydroxylase; oeNC: over-expression negtive control; 6-OHDA: 6-hydroxydopamine, 100 μmol/L; MG: mangiferin, 50 μmol/L, one-way ANOVA.

Article Snippet: Primary antibodies are listed as follow: TH (1:5000, 25859-1-AP, proteintech), α-synuclein (1:1000, ab212184, Abcam), phosphate- GSK-3β (1:1000, AF2016, Affinity), GSK-3β (1:2000, ab93926, Abcam), AKR1C3 (1:2000, 11194-1-AP, proteintech), wnt1 (1:2000, 27935-1-AP, proteintech), β-catenin (1:10000, 51067-2-AP, proteintech), caspase-3 (1:2000, ab184787, Abcam), GAPDH (1:10000, ab8245, Abcam).

Techniques: Activation Assay, Gene Expression, Quantitative RT-PCR, Expressing, Western Blot, Control, Over Expression

Journal: Neuroscience letters

Article Title: Mangiferin alleviates 6-OHDA-induced Parkinson's disease by inhibiting AKR1C3 to activate Wnt signaling pathway.

doi: 10.1016/j.neulet.2023.137608

Figure Lengend Snippet: Fig. 7. Attenuation of mangiferin's improvement on motor behavior in PD mice with AKR1C3 activation in vivo. (A) Flowchart depicting the establishment of the PD mouse model, drug treatment, behavioral testing, and biological evaluation. day 0 was considered the day when stereotaxic injections of lentivirus or lentiviral empty vector were administered to the lesion side of the striatum, and mice were raised for three weeks. From days 14 to 28, Mangiferin was administered i.p. at a dose of 30mg/kg/day for 14 days, and behavioral training and experiments were conducted from days 21 to 28. At the end of the experiment, animals were euthanized with excess isoflurane anesthesia. The substantia nigra was extracted for the measurement of MDA and SOD levels via ELISA, and WB and RT-qPCR was applied. (B) Impact of mangiferin on fatigue stick-turning time in PD mouse model. (C) Effects of mangiferin on pole-

Article Snippet: Primary antibodies are listed as follow: TH (1:5000, 25859-1-AP, proteintech), α-synuclein (1:1000, ab212184, Abcam), phosphate- GSK-3β (1:1000, AF2016, Affinity), GSK-3β (1:2000, ab93926, Abcam), AKR1C3 (1:2000, 11194-1-AP, proteintech), wnt1 (1:2000, 27935-1-AP, proteintech), β-catenin (1:10000, 51067-2-AP, proteintech), caspase-3 (1:2000, ab184787, Abcam), GAPDH (1:10000, ab8245, Abcam).

Techniques: Activation Assay, In Vivo, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Neuroscience letters

Article Title: Mangiferin alleviates 6-OHDA-induced Parkinson's disease by inhibiting AKR1C3 to activate Wnt signaling pathway.

doi: 10.1016/j.neulet.2023.137608

Figure Lengend Snippet: Fig. 8. Attenuation of mangiferin's protective effect in PD with AKR1C3 activation in vivo. (A) The impact of AKR1C3 activation on MDA and SOD levels in vivo. (B) The effects of AKR1C3 activation on TH protein and α-synuclein protein in vivo. *P<0.05, **P<0.01, ***P<0.001 vs the control group, #P<0.05, ##P<0.01, 6- OHDA+MG (50 μM) group vs 6-OHDA group, n=6. PD: Parkinson’s disease; oeNC: over-expression negtive control; TH: tyrosine hydroxylase; 6-OHDA: 6- hydroxydopamine, 100μmol/L; MG: mangiferin, 50μmol/L, one-way ANOVA.

Article Snippet: Primary antibodies are listed as follow: TH (1:5000, 25859-1-AP, proteintech), α-synuclein (1:1000, ab212184, Abcam), phosphate- GSK-3β (1:1000, AF2016, Affinity), GSK-3β (1:2000, ab93926, Abcam), AKR1C3 (1:2000, 11194-1-AP, proteintech), wnt1 (1:2000, 27935-1-AP, proteintech), β-catenin (1:10000, 51067-2-AP, proteintech), caspase-3 (1:2000, ab184787, Abcam), GAPDH (1:10000, ab8245, Abcam).

Techniques: Activation Assay, In Vivo, Control, Over Expression

Journal: Neuroscience letters

Article Title: Mangiferin alleviates 6-OHDA-induced Parkinson's disease by inhibiting AKR1C3 to activate Wnt signaling pathway.

doi: 10.1016/j.neulet.2023.137608

Figure Lengend Snippet: Fig. 9. Suppression of Wnt signaling activation by AKR1C3 overexpression in vivo effects of mangiferin. (A) Measurement of gene expression levels of GSK-3β, Wnt1, and β-catenin in AKR1C3 overexpressing cells using RT-qPCR. (B) Analysis of protein expression levels of p-GSK-3β, GSK-3β, Wnt1, and β-catenin through Western blotting in AKR1C3 overexpressing cells. *P<0.05, **P<0.01, ***P<0.001 vs the control group, #P<0.05, ##P<0.01, ###P<0.001, 6-OHDA+MG(50μM) group vs 6-OHDA group, n=6. oeNC: over-expression negtive control; 6-OHDA: 6-hydroxydopamine, 100 μmol/L; MG: mangiferin, 50 μmol/L, one-way ANOVA.

Article Snippet: Primary antibodies are listed as follow: TH (1:5000, 25859-1-AP, proteintech), α-synuclein (1:1000, ab212184, Abcam), phosphate- GSK-3β (1:1000, AF2016, Affinity), GSK-3β (1:2000, ab93926, Abcam), AKR1C3 (1:2000, 11194-1-AP, proteintech), wnt1 (1:2000, 27935-1-AP, proteintech), β-catenin (1:10000, 51067-2-AP, proteintech), caspase-3 (1:2000, ab184787, Abcam), GAPDH (1:10000, ab8245, Abcam).

Techniques: Activation Assay, Over Expression, In Vivo, Gene Expression, Quantitative RT-PCR, Expressing, Western Blot, Control

Journal: Sexual development : genetics, molecular biology, evolution, endocrinology, embryology, and pathology of sex determination and differentiation

Article Title: Testosterone synthesis in patients with 17β-hydroxysteroid dehydrogenase 3 deficiency.

doi: 10.1159/000336605

Figure Lengend Snippet: Fig. 6. Immunohistochemical localization of AKR1C3 in the testis of patient ARD1853 ( a ) and a normal control ( b ). Leydig cells (white arrowheads) and peritubular cells (yellow arrowheads) show a strong expression of AKR1C3.

Article Snippet: Sections were immunostained with goat anti-AKR1C3 antibody (NB-100–1940, Novus) diluted 1: 160.

Techniques: Immunohistochemical staining, Control, Expressing

Journal: Molecular Cancer Therapeutics

Article Title: Resistance to Pyrrolobenzodiazepine Dimers Is Associated with SLFN11 Downregulation and Can Be Reversed through Inhibition of ATR

doi: 10.1158/1535-7163.mct-20-0351

Figure Lengend Snippet: Figure 2. Proteomic profiling of 361-PBDr cells identified differentially expressed proteins. A, Experimental workflow of the quantitative proteomic profiling. Equal numbers of cells were lysed from duplicate samples of two cell lines. Extracted proteins was digested into tryptic peptides, which were labeled by TMT and analyzed by HPLC-MS/MS. Proteins were subsequently identified and quantified. B, Cellular localization analysis of the identified proteins. C, Volcano plot summarizing protein quantification results of 361-PBDr cells compared with parental cells. Shaded regions indicate DEPs with log2 fold change ≥0.45 or ≤0.45 and significance P < 0.05. D, Quantitative RT-PCR assessed the differential RNA expression in 361-PBDr cells compared with parental cells. Relative mRNA abundance are represented as fold change in 361-PBDr cells compared with parental cells SD from two experiments. E, Western blot analysis demonstrated the differential protein expression of SLFN11, S100A4, AKR1C3, and galectin-1 in resistant 361-PBDr cells compared with parental MDA-MB-361 cells.

Article Snippet: Membranes were probed with SLFN11 (D2) and CHK1 antibody (Santa Cruz Biotechnology) at 1:100 dilution, orwith S100A4, galectin-1, pCHK1(S345), andb-actin antibodies (Cell Signaling Technology) at 1:1,000 dilutions, or with AKR1C3 antibody (R & D Systems) at 1:500 dilution.

Techniques: Labeling, Tandem Mass Spectroscopy, Quantitative RT-PCR, RNA Expression, Western Blot, Expressing