a 3 ar agonists  (BioIntervene Inc)

Journal: Antioxidants

Article Title: Characterization of the Peroxisomal Proteome and Redox Balance in Human Prostate Cancer Cell Lines

doi: 10.3390/antiox13111340

Figure Lengend Snippet: Figure 9. Treatment of LNCaP cells with 10 nM R1881 modulates the peroxisomal glutathione redox state and H2O2 levels. LNCaP cells were cultured in phenol red-free MEM supplemented with 10% charcoal-stripped serum for 4 days. Subsequently, they were treated with either the ethanol vehicle (−) or 1 pM or 10 nM R1881 (+). After one day, the cells were transfected with a plasmid encoding a peroxisomal (PO), cytosolic (C), or mitochondrial (MT) variant of (A) the glutathione redox sensor roGFP2, or (B) the H2O2 sensor roGFP2-Orp1 (representative images of the subcellular distribution patterns of the fluorescent reporter proteins are shown in Figure S8). Subsequently, the medium was replaced with fresh ethanol- or R1881-containing medium. Two days later, the F400/F480 response ratios of the sensors were measured, expressed as the percentage of the average vehicle response, and presented as box plots. Each box represents the interquartile range, with the bottom and top showing the 25th and 75th percentiles, respectively. The line inside the box indicates the median and the lines extending from the box representing one standard deviation below and above the mean. Data from each independent experiment (n = 3) are indicated by a different color. Statistical comparisons were performed using nested T-tests (ns, non-significant; *, p < 0.05; **, p < 0.01; ****, p < 0.0001). AU, arbitrary units.

Article Snippet: In experiments involving LNCaP cells treated with the AR agonist R1881 (E3164-000, Steraloids, Newport, RI, USA), the cells were cultured in phenol red-free MEM (51200046, Gibco, Billings, MT, USA) supplemented with 10% charcoal-stripped FBS (S181F-100, Avantor), 2 mM ultraglutamine-1, and 0.2% (v/v) Mycozap.

Techniques: Cell Culture, Transfection, Plasmid Preparation, Variant Assay, Standard Deviation

Journal: Antioxidants

Article Title: Characterization of the Peroxisomal Proteome and Redox Balance in Human Prostate Cancer Cell Lines

doi: 10.3390/antiox13111340

Figure Lengend Snippet: Figure 10. Gene-set enrichment analysis of differentially expressed proteins in LNCaP cells treated with the AR agonist R1881 compared to the vehicle. LNCaP cells were cultured in phenol red-free MEM supplemented with 10% charcoal-stripped serum for 4 days, followed by treatment with either ethanol (vehicle) or 10 nM R1881. After one day, the medium was replaced with fresh ethanol- or R1881-containing medium. Two days later, cells were harvested and processed for proteomics analysis. (A) Volcano plots showing the differences in protein abundance between 10 nM R1881-treated and vehicle-treated LNCaP cells. Proteins significantly up- or downregulated (FC ≥2; pAdj < 0.05) are indicated by red or blue dots, respectively (n = 3 biological replicates). Black dots represent proteins with no significant difference. Numbers within the panel denote the total number of proteins in each group. The list of differentially expressed proteins was generated using FragPipe-Analyst, and the Benjamin–Hochberg method was applied to correct for the FDR. (B,C) Molecular Signatures Database (MsigDB) hallmarks and KEGG enrichment analyses of the differentially expressed proteins were conducted using ShinyGO 0.80. Settings included selected species (human), FDR cut-off (0.05), and sorting by FDR. The total list of identified proteins was uploaded as the background. KEGG pathways and MsigDB hallmarks with significant enrichment (up to 10) are shown for the (B) upregulated and (C) downregulated differentially expressed proteins, sorted by fold enrichment. Circle size represents the number of proteins identified in each pathway/hallmark. DN, down-regulated; E2F, transcription factor E2F; IL2, interleukin 2; NFKB, nuclear factor kappa-B; PPAR, peroxisome proliferator-activated receptor; STAT5, signal transducer and activator of transcription 5; TNFA, tumor necrosis factor-alpha; UV, ultraviolet.

Article Snippet: In experiments involving LNCaP cells treated with the AR agonist R1881 (E3164-000, Steraloids, Newport, RI, USA), the cells were cultured in phenol red-free MEM (51200046, Gibco, Billings, MT, USA) supplemented with 10% charcoal-stripped FBS (S181F-100, Avantor), 2 mM ultraglutamine-1, and 0.2% (v/v) Mycozap.

Techniques: Cell Culture, Quantitative Proteomics, Generated

Journal: Antioxidants

Article Title: Characterization of the Peroxisomal Proteome and Redox Balance in Human Prostate Cancer Cell Lines

doi: 10.3390/antiox13111340

Figure Lengend Snippet: Figure 11. Heatmaps depicting the proteome profile of 72 peroxisome-associated proteins in LNCaP cells treated with the AR agonist R1881 relative to the vehicle. The relative protein abundances, expressed as Log2 FC, are shown on a false color scale with red for upregulation, white for equal expression, and blue for downregulation compared to vehicle-treated LNCaP cells. The cells were cultured in phenol red-free MEM supplemented with 10% (v/v) charcoal-stripped serum for 4 days, followed by treatment with either ethanol (vehicle) or 10 nM R1881. After one day, the medium was replaced with fresh ethanol- or R1881-containing medium. Two days later, cells were harvested and processed for proteomics analysis. Each box represents the average of 3 biological replicates, with Log2-FC shown only if the adjusted p-value is <0.05.

Article Snippet: In experiments involving LNCaP cells treated with the AR agonist R1881 (E3164-000, Steraloids, Newport, RI, USA), the cells were cultured in phenol red-free MEM (51200046, Gibco, Billings, MT, USA) supplemented with 10% charcoal-stripped FBS (S181F-100, Avantor), 2 mM ultraglutamine-1, and 0.2% (v/v) Mycozap.

Techniques: Expressing, Cell Culture

Journal: Antioxidants

Article Title: Characterization of the Peroxisomal Proteome and Redox Balance in Human Prostate Cancer Cell Lines

doi: 10.3390/antiox13111340

Figure Lengend Snippet: Figure 12. The AR agonist R1881 decreases catalase activity in LNCaP cells. LNCaP cells were cultured in phenol red-free MEM supplemented with 10% (v/v) charcoal-stripped serum for 4 days. The cells were then treated with either ethanol (vehicle) or 10 nM R1881. After one day, the medium was replaced with fresh medium containing ethanol, or R1881. Two days later, CAT activity was measured (n = 4). Error bars represent the standard deviation. Statistical significance was assessed using an unpaired T-test (**, p < 0.01).

Article Snippet: In experiments involving LNCaP cells treated with the AR agonist R1881 (E3164-000, Steraloids, Newport, RI, USA), the cells were cultured in phenol red-free MEM (51200046, Gibco, Billings, MT, USA) supplemented with 10% charcoal-stripped FBS (S181F-100, Avantor), 2 mM ultraglutamine-1, and 0.2% (v/v) Mycozap.

Techniques: Activity Assay, Cell Culture, Standard Deviation

Journal: Antioxidants

Article Title: Characterization of the Peroxisomal Proteome and Redox Balance in Human Prostate Cancer Cell Lines

doi: 10.3390/antiox13111340

Figure Lengend Snippet: Figure 13. Heatmaps showing the proteome profile of 36 primary and secondary antioxidant enzymes in LNCaP cells treated with the AR agonist R1881 compared to the vehicle. The cells were cultured in phenol red-free MEM supplemented with 10% (v/v) charcoal-stripped serum for 4 days. The cells were then treated with either ethanol (vehicle) or 10 mM R1881. After one day, the medium was replaced with fresh medium containing either ethanol or R1881. Two days later, cells were harvested and processed for proteomics analysis. Each box represents the average of 3 biological replicates, with Log2 FC shown only if the adjusted p-value is <0.05.

Article Snippet: In experiments involving LNCaP cells treated with the AR agonist R1881 (E3164-000, Steraloids, Newport, RI, USA), the cells were cultured in phenol red-free MEM (51200046, Gibco, Billings, MT, USA) supplemented with 10% charcoal-stripped FBS (S181F-100, Avantor), 2 mM ultraglutamine-1, and 0.2% (v/v) Mycozap.

Techniques: Cell Culture