Journal: International Journal of Neuropsychopharmacology

Article Title: A Characterization of the Effects of Minocycline Treatment During Adolescence on Structural, Metabolic, and Oxidative Stress Parameters in a Maternal Immune Stimulation Model of Neurodevelopmental Brain Disorders

doi: 10.1093/ijnp/pyab036

Figure Lengend Snippet: Minocycline-Related Effects on the Expression of Inflammatory and Oxidative Markers in VH and MIS Offspring

Article Snippet: Blots were blocked with 5% bovine serum albumin (BSA) (Sigma, Spain) for 1 hour at room temperature and probed overnight at 4°C with rabbit anti-iNOS (sc-650, 1:750 BSA 2%; SCBT), goat anti-COX2 (sc-1747, 1:750 BSA 2.5%; SCBT), mouse anti-KEAP1 (MAB3024, 1:1000; R&D), rabbit anti-HO1 (ab68477, 1:1000; Abcam), goat anti-NQO1 (sc16464, 1:750; BSA 1%, SCBT), and mouse anti-β-actin (A5441, 1:10 000; Sigma).

Techniques: Expressing

Journal: International Journal of Neuropsychopharmacology

Article Title: A Characterization of the Effects of Minocycline Treatment During Adolescence on Structural, Metabolic, and Oxidative Stress Parameters in a Maternal Immune Stimulation Model of Neurodevelopmental Brain Disorders

doi: 10.1093/ijnp/pyab036

Figure Lengend Snippet: Minocycline-related effects on the expression of IOS markers in VH and MIS animals. Inflammatory markers expression (iNOS, COX2), antioxidant enzyme activity (GPx, CAT, SOD), and oxidative stress markers (MDA, NRF2, KEAP1, HO1, NQO1, GSH free , GSH total , GSSG) in prefrontal cortex, hippocampus, caudate-putamen, and amygdala (n = 7–8 animals). Representative bands of iNOS, COX2, KEAP1, HO1, and NQO1 (upper bands) and the loading control, β-actin (lower bands), are shown above their corresponding graph bars. Each column represents the mean ± SEM of 5–8 animals. Two-way ANOVA followed by a pairwise interaction contrast (* P < .05, ** P < .01, *** P < .001, vs Sal-treated animals, & P < .05, && P < .01, &&& P < .001 vs VH animals).

Article Snippet: Blots were blocked with 5% bovine serum albumin (BSA) (Sigma, Spain) for 1 hour at room temperature and probed overnight at 4°C with rabbit anti-iNOS (sc-650, 1:750 BSA 2%; SCBT), goat anti-COX2 (sc-1747, 1:750 BSA 2.5%; SCBT), mouse anti-KEAP1 (MAB3024, 1:1000; R&D), rabbit anti-HO1 (ab68477, 1:1000; Abcam), goat anti-NQO1 (sc16464, 1:750; BSA 1%, SCBT), and mouse anti-β-actin (A5441, 1:10 000; Sigma).

Techniques: Expressing, Activity Assay

Journal: Cell Death & Disease

Article Title: Blockage of Nrf2 and autophagy by L-selenocystine induces selective death in Nrf2-addicted colorectal cancer cells through p62-Keap-1-Nrf2 axis

doi: 10.1038/s41419-022-05512-2

Figure Lengend Snippet: A – C Translocation of phosphorylated Nrf2 into the nucleus in non–Nrf2-addicted MSCs ( A ) and WiDr ( B ) and C 2 BBe 1 ( C ) cells. The cell count in field was determined using Image J. The fluorescence intensity was normalized with cell number from eight field. All quantified results are express as mean ± SD. (* P < 0.05, ** P < 0.01, *** P < 0.001, compared with solvent control for each cell type). D Protein levels of NRF2, Keap1, p62, NQO1, and xCT in MSCs and WiDr and C 2 BBe 1 cells after 24 h of SeC treatment. Relative levels of these target proteins were normalized to GAPDH (loading control. The values were presented as average ± SD from three independent repeats. (* and + P < 0.05, ** and ++ P < 0.01, * ** and +++ P < 0.001, n = 3, compared with solvent control for each cell type). Significance was determined by an unpaired t test.

Article Snippet: NFE2L2 human siRNA oligo duplex and universal scrambled negative control duplex were obtained from OriGene. siGENOME human KEAP1 siRNA-smart pool and nontarget siRNA pool were purchased from Dharmacon siGENOME.

Techniques: Translocation Assay, Cell Counting, Fluorescence

Journal: Cell Death & Disease

Article Title: Blockage of Nrf2 and autophagy by L-selenocystine induces selective death in Nrf2-addicted colorectal cancer cells through p62-Keap-1-Nrf2 axis

doi: 10.1038/s41419-022-05512-2

Figure Lengend Snippet: A Nrf2, Keap1, p62, NQO1, HO1, xCT, ULK1, Beclin-1, and LC3II levels in C 2 BBe 1 cells after SeC treatment for 48 h (* p < 0.05, * p < 0.01, *** p < 0.001, compared with solvent control, significance was determined by an unpaired t test). The values were presented as average ± SD from three independent repeats. B Relative viability of SeC-treated C 2 BBe 1 cells with or without GSH pretreatment for 48 h. The values were presented as average ± SD from eight independent repeats. (* p < 0.05, ** p < 0.01, *** p < 0.001, compared with solvent control; ## p < 0.01, ### p < 0.001, compared with SeC alone, significance was determined by one-way ANOVA).

Article Snippet: NFE2L2 human siRNA oligo duplex and universal scrambled negative control duplex were obtained from OriGene. siGENOME human KEAP1 siRNA-smart pool and nontarget siRNA pool were purchased from Dharmacon siGENOME.

Techniques:

Journal: Cell Death & Disease

Article Title: Blockage of Nrf2 and autophagy by L-selenocystine induces selective death in Nrf2-addicted colorectal cancer cells through p62-Keap-1-Nrf2 axis

doi: 10.1038/s41419-022-05512-2

Figure Lengend Snippet: A siRNA targeting scramble control (scr) and Nrf2 transfected into WiDr cells to alleviate their Nrf2 addiction status. The relative viability determined using the MTT assay ( n = 5). The values were presented as average ± SD from five independent repeats. B siRNA targeting scr and Keap1 transfected into C 2 BBe 1 cells to increase Nrf2 addiction status. The relative viability was determined using the MTT assay ( n = 4). The ROS levels was measured after 1 h of SeC treatment ( n = 5). The values were presented as average ± SD from four or five independent repeats. (* p < 0.05, * p < 0.01, *** p < 0.001, compared with scr). C Persist treatment with 50 μM tBHQ for 2 weeks to mimic Nrf2 overactivation in C 2 BBe 1 cells. The relative viability was determined after SeC treatment for 48 h using the MTT assay ( n = 8). The values were presented as average ± SD from eight independent repeats. (* p < 0.05, ** p < 0.01, *** p < 0.001, compared with solvent control; ## p < 0.01, ### p < 0.001, compared with SeC alone). Significance was determined by two-way ANOVA.

Article Snippet: NFE2L2 human siRNA oligo duplex and universal scrambled negative control duplex were obtained from OriGene. siGENOME human KEAP1 siRNA-smart pool and nontarget siRNA pool were purchased from Dharmacon siGENOME.

Techniques: Transfection, MTT Assay