Journal: Redox Biology

Article Title: Female sex is a risk factor for exacerbated lipid peroxidation and disease in murine retinitis pigmentosa

doi: 10.1016/j.redox.2025.103987

Figure Lengend Snippet: Significantly elevated retinal lipid peroxidation is evident in adult P23H female mice but not in P23H male mice. The figure presents normalized expression of A-F) oxidative stress markers, G-J) genes encoding reactive oxygen species (ROS) clearing enzymes, K-M) superoxide dismutase 2 (SOD2) and catalase (CAT) protein expression, and N) 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity (a measure of antioxidant efficiency) across age and sex in P23H and WT mouse whole retinal extracts, mean ± SD. A) 4-hydroxynonenal (4-HNE) content as measured with an enzyme-linked immunosorbent assay (ELISA), fold-change (FC) to WT. B) 4-HNE expression as measured from antibody staining intensity in the immunohistochemistry (IHC) assay, FC compared to WT. C) Upper section: Representative raw images of 4-HNE signal used in the analysis with the corresponding negative control image from 5-month-old retinal cryosections - Lower section: Corresponding magnified IHC images as obtained from 5-month-old retinal cryosections stained with a 4-HNE antibody (green) and counterstained with the nuclei marker 4′,6-diamidino-2-phenylindole (DAPI; blue). D-E) Protein carbonyl content as measured with D) ELISA, or E) immunoblotting/western blotting (WB), FC compared to WT. F) A representative carbonyl immunoblot image presenting data obtained from 5-month-old mouse retinas. G-J) Expression of genes encoding ROS clearing enzymes G) Sod1 ; H) Sod2 ; I) Cat ; and J) glutathione peroxidase 4 ( Gpx4 ), as measured with quantitative polymerase chain reaction (qPCR), log10(2 -(ΔΔCt) ). K) Example images from SOD2 and CAT immunoblotting. L) SOD2 expression as measured with WB, FC to WT. M) CAT expression as measured with immunoblotting, FC to WT. N) DPPH scavenging activity, FC to WT. The asterisks indicate significant one-way ANOVA posthoc (Tukey's) test results, ∗ = p < 0.05, ∗∗ = p < 0.01 and ∗∗∗ = p < 0.001.

Article Snippet: 4-HNE ELISA (E-EL-0128, Elabscience) was conducted according to the manufacturer's instructions.

Techniques: Expressing, Activity Assay, Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemistry, Negative Control, Marker, Western Blot, Real-time Polymerase Chain Reaction

Journal: Redox Biology

Article Title: Female sex is a risk factor for exacerbated lipid peroxidation and disease in murine retinitis pigmentosa

doi: 10.1016/j.redox.2025.103987

Figure Lengend Snippet: Investigation of inflammation and oxidative stress in rd10 mouse retinas reveals higher lipid peroxidation in 2-month-old rd10 females, whereas inflammation and antioxidant capacity show only disease-dependent changes. A) Normalized expression of glial fibrillary acidic protein (GFAP) in the immunoblotting/western blotting (WB) assay, fold-change (FC) compared to WT. B) Representative immunoblots as obtained from 2- and 4 -month-old WT and rd10 retina extracts. C) 4-hydroxynonenal (4-HNE) content as measured with an enzyme-linked immunosorbent assay (ELISA), FC compared to WT. D) Protein carbonyl content measured with ELISA, FC compared to WT. E) Antioxidative capacity to scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH), FC to WT. Data is presented as mean ± SD, and the asterisks mark significant one-way ANOVA posthoc (Tukey's) test results, ∗ = p < 0.05, ∗∗ = p < 0.01, ∗∗∗ = p < 0.001 and ∗∗∗∗ = p < 0.0001.

Article Snippet: 4-HNE ELISA (E-EL-0128, Elabscience) was conducted according to the manufacturer's instructions.

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Redox Biology

Article Title: Female sex is a risk factor for exacerbated lipid peroxidation and disease in murine retinitis pigmentosa

doi: 10.1016/j.redox.2025.103987

Figure Lengend Snippet: WT mice display no sex differences in retinal function, thickness, GFAP expression or lipid peroxidation. A) Scotopic and B) photopic ERG amplitudes (μV) in WT female versus male mice. WT 2-MO, n = 8 males (m), n = 8 females (f) in scotopic ERG, and n = 3 m, n = 3 f in photopic ERG. WT 5-MO, n = 5 m, n = 5 f. C) Mean outer nuclear layer (ONL) thickness (μm) from four retinal orientations (superior, inferior, nasal, temporal) in 2- and 5-MO WT mice. D) Normalized expression of glial fibrillary acidic protein (GFAP) in the immunoblotting/western blotting (WB) assay in post-pubertal (=2–5 months of age) WT mice, fold-change (FC) compared to male WT. E) 4-hydroxynonenal (4-HNE) content as measured with an enzyme-linked immunosorbent assay (ELISA) in pre- (1-month-old) and post-pubertal WT mice, fold-change (FC) to male WT. F) 4-HNE expression as measured from antibody staining intensity in the immunohistochemistry (IHC) assay in post-pubertal WT mice, FC compared to WT male group. Data is presented as mean ± SD.

Article Snippet: 4-HNE ELISA (E-EL-0128, Elabscience) was conducted according to the manufacturer's instructions.

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemistry