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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 <t>(SOD2)</t> 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.
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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 <t>(SOD2)</t> 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.
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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 <t>(SOD2)</t> 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.
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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 <t>(SOD2)</t> 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.
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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 <t>(SOD2)</t> 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.
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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 <t>(SOD2)</t> 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.
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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.

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: After 3 washes with TBST, the membranes were incubated overnight in primary antibody (anti-mouse GFAP 1:2000 (3670, Cell Signaling Technology), anti-rabbit SOD2 1:5000 (ab13533, Abcam), anti-mouse CAT 1:500 (sc-271803, Santa Cruz Biotechnology)) in 5 % milk TBST.

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