Journal: Frontiers in Medicine

Article Title: Restored retinal physiology after administration of niacin with citicoline in a mouse model of hypertensive glaucoma

doi: 10.3389/fmed.2023.1230941

Figure Lengend Snippet: Western blot antibodies.

Article Snippet: Rabbit polyclonal anti-Nrf-2 , 1:1000 , Abcam (Cambridge, UK) , ab92946.

Techniques: Western Blot

Journal: Frontiers in Medicine

Article Title: Restored retinal physiology after administration of niacin with citicoline in a mouse model of hypertensive glaucoma

doi: 10.3389/fmed.2023.1230941

Figure Lengend Snippet: Effect of individual or combined administration of niacin and citicoline on MCE-induced oxidative stress. (A) Representative Western blots and densitometric analysis of nuclear factor erythroid 2-related factor-2 (Nrf-2) and (B) heme oxygenase-1 (HO-1) levels in control and MCE mice either untreated or treated with individual niacin and citicoline or their combination. Data are expressed as mean ± SEM ( n = 6 retinas for group). * p < 0.0001 vs. control; § p < 0.01, §§ p < 0.001, and §§§ p < 0.0001 vs. MCE (two-way ANOVA followed by Tukey’s multiple comparison post-hoc test).

Article Snippet: Rabbit polyclonal anti-Nrf-2 , 1:1000 , Abcam (Cambridge, UK) , ab92946.

Techniques: Western Blot, Comparison

Journal: Antioxidants

Article Title: Evidence for TGF-β1/Nrf2 Signaling Crosstalk in a Cuprizone Model of Multiple Sclerosis

doi: 10.3390/antiox13080914

Figure Lengend Snippet: TGF-β1 receptor 1 blocking reduces both Nrf2 and TGF-β1 protein levels in the CPZ model of MS. ( A , B ) Protein levels in respect to the control and normalized by β-actin for Nrf2 ( A ) and TGF-β1 ( B ) in the corpus callosum of the mice fed with CPZ over 3 or 5 weeks (3W and 5W) treated with galunisertib (GAL, 10 mg/kg, red blocks) or vehicle (Veh, gray blocks). Representative images of western blot membranes from two mice are shown (bottom). Kruskal–Wallis test, Dunn’s multiple comparison post hoc tests (*) p > 0.05, (**), p < 0.01. ( n = 4–8 mice). ( C ) Correlation between protein levels for TGF-β1 and Nrf2 at the different time points and treatments (r = 0.835, p < 0.001, Spearman test).

Article Snippet: The reagents and antibodies used were as follows: Cuprizone (C9012-25G), DMSO, ketamine, xylazine, paraformaldehyde (P6148-500G) (from Sigma-Aldrich, Darmstadt, Germany), Galunisertib (GAL; HY-13226, MedchemExpress, Princeton, NJ, USA), normal goat serum (NGS; 50062Z, Life Technologies, Carlsbad, CA, USA), 0.5% Triton X-100 (E5111) and mouse anti-CC1 antibody to detect oligodendrocytes (MA1 25884; 1:300) (Thermo Fisher Scientific, Waltham, MA, USA), chicken anti-myelin basic protein to detect myelin (MBP; 1:800; Invitrogen, Waltham MA, USA), rabbit anti-phosphorylated Smad2 (138D4; 1:1000), anti-Smad 2/3 (D7G7, 1:1000), rabbit anti-Nrf2 (D1Z96; 1:1000 WB, 1:200 IF) (Cell Signaling, Danvers, MA, USA); mouse anti-TGF-β1 (sc-130348; 1:800); and rabbit anti-Iba1 (sc-32725; 1:50) (from Santa Cruz, TX, USA).

Techniques: Blocking Assay, Control, Western Blot, Comparison

Journal: Antioxidants

Article Title: Evidence for TGF-β1/Nrf2 Signaling Crosstalk in a Cuprizone Model of Multiple Sclerosis

doi: 10.3390/antiox13080914

Figure Lengend Snippet: Putative mechanisms of the TGF-β1/Nrf2 signaling crosstalk in demyelinated lesions. Under demyelination by cuprizone, TGF-β1 expression increases, most likely promoting astrocyte and microglia TGF-β1 release. The activation of the TGF-β1 receptor will, in turn, activate an Nrf2-dependent antioxidant response, such as enhanced catalase (CAT) expression and ATP synthesis. Thus, blocking the TGF-β1 receptor reduces this response by mechanisms to be determined (?), triggering a failure in antioxidant-related recovery (for instance, locomotor and cognitive function). This figure was created in Biorender.

Article Snippet: The reagents and antibodies used were as follows: Cuprizone (C9012-25G), DMSO, ketamine, xylazine, paraformaldehyde (P6148-500G) (from Sigma-Aldrich, Darmstadt, Germany), Galunisertib (GAL; HY-13226, MedchemExpress, Princeton, NJ, USA), normal goat serum (NGS; 50062Z, Life Technologies, Carlsbad, CA, USA), 0.5% Triton X-100 (E5111) and mouse anti-CC1 antibody to detect oligodendrocytes (MA1 25884; 1:300) (Thermo Fisher Scientific, Waltham, MA, USA), chicken anti-myelin basic protein to detect myelin (MBP; 1:800; Invitrogen, Waltham MA, USA), rabbit anti-phosphorylated Smad2 (138D4; 1:1000), anti-Smad 2/3 (D7G7, 1:1000), rabbit anti-Nrf2 (D1Z96; 1:1000 WB, 1:200 IF) (Cell Signaling, Danvers, MA, USA); mouse anti-TGF-β1 (sc-130348; 1:800); and rabbit anti-Iba1 (sc-32725; 1:50) (from Santa Cruz, TX, USA).

Techniques: Expressing, Activation Assay, Blocking Assay

Journal: Cells

Article Title: Lipoic Acid Synergizes with Antineoplastic Drugs in Colorectal Cancer by Targeting p53 for Proteasomal Degradation

doi: 10.3390/cells8080794

Figure Lengend Snippet: LA induces Nrf2, which is likely dispensable for p53 degradation. ( A ) HCT116 cells were incubated with increasing doses of LA (125–1000 µM) for 48 h. Cells were thereafter subjected to SDS-PAGE and western blot analysis of p53, Nrf2 and its downstream target HO-1. EtOH (0 µM) served as solvent control. Hsp90 was visualized as loading control. ( B ) HCT116 cells were incubated for 48 h with LA in the presence or the absence of ML385, a pharmacological Nrf2 inhibitor. Hemin (200 µM, 24 h) was included as positive control for HO-1 induction. EtOH (0 µM) served as vehicle control. Cells were then lyzed and underwent western blot analysis of Nrf2, p53, HO-1, as well as p62. Hsp90 was used as loading control. ( C – F ) Densitometric quantification of NRF2 (C), p53 (D), HO-1 (E) and p62 (F) obtained from three independent experiments as described and shown in B. Data are given as mean + SEM ( n = 3). ns p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Primary antibodies included Hsp90α/β (F8, mouse monoclonal; Santa Cruz, no. sc-13119), p53 (DO-1, mouse monoclonal; Santa Cruz, no. sc-126), p53 (FL-393; rabbit polyclonal; Santa Cruz, no. sc-6243), p62 (mouse monoclonal; Santa Cruz, no. sc-28359), LC3B (rabbit monoclonal; Cell Signaling Technology, no. 3868), ATG5 (rabbit monoclonal, Cell Signaling Technology, no. 12994), ubiquitin (mouse monoclonal; Santa Cruz, no. sc-8017), Nrf2 antibody (rabbit monoclonal; GeneTex, no. GTX103322), MDM2 (mouse monoclonal; Santa Cruz, no. sc-56154), heme oxygenase-1 (HO-1; rabbit polyclonal; GeneTex, no. GTX101147), as well as p21 (C-19, rabbit polyclonal; Santa Cruz, no. sc-397).

Techniques: Incubation, SDS Page, Western Blot, Positive Control