Journal: Biology of Sex Differences

Article Title: Sex differences in ferroptosis-related vulnerability to autism-like deficits in the adolescent medial prefrontal cortex following embryonic valproic acid exposure

doi: 10.1186/s13293-026-00982-x

Figure Lengend Snippet: E12.5 VPA exposed induces male-biased ferroptosis-associated alterations in the mPFC. A Representative protein bands of GPX4, SLC7A11/xCT, ACSL4 and FTH1 in mPFC. B-E Statistical graph of protein levels of GPX4, SLC7A11/xCT, ACSL4 and FTH1. n = 5 per group. F Content of ferrous iron in the P24 mPFC. n = 3 per group. G GSH concentration and the ratio of GSH and GSSH in the P24 mPFC measured. n = 3 per group. H The MDA level in the P24 mPFC. n = 3 per group. I The SOD concentration in the P24 mPFC. n = 3 per group. J Representative immunofluorescence images of SLC7A11 (red) and DAPI (blue) in mPFC. Scale bar = 100 μm. Mean fluorescence intensity of SLC7A11/xCT, n = 3 per group. K Representative immunofluorescence images of ACSL4 (red) and DAPI (blue) in mPFC. Scale bar = 100 μm. Mean fluorescence intensity of ACSL4, n = 3 per group. L Representative immunofluorescence images of ROS (red) and DAPI (blue) in mPFC. Scale bar = 100 μm. Mean fluorescence intensity of ACSL4, n = 3 per group. M Electron microscope of mitochondrial morphology in the P24 mPFC. The second row represents a partial enlargement of the first row, respectively. Scale bar = 2 μm (raw) / 500 nm (zoom). Values was presented as mean ± SEM and analyzed by one-way ANOVA. ns > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The membranes were then incubated overnight at 4 °C with the following primary antibodies: GPX4 (1:2000; Cat# A11243; Abclonal), FTH1 (1:5000; Cat# A19544; Abclonal), ACSL4 (1:1000; Cat#38493T; Cell Signaling Technology), xCT/SLC7A11 (1:1000; Cat# 28001T; Cell Signaling Technology), and β-actin (1:50000; Cat# 66009-1-Ig; Proteintech).

Techniques: Concentration Assay, Immunofluorescence, Fluorescence, Microscopy

Journal: Biology of Sex Differences

Article Title: Sex differences in ferroptosis-related vulnerability to autism-like deficits in the adolescent medial prefrontal cortex following embryonic valproic acid exposure

doi: 10.1186/s13293-026-00982-x

Figure Lengend Snippet: E12.5 VPA induces ferroptosis-related signatures in the hippocampus without sex differences. A Representative protein bands of GPX4, SLC7A11/xCT, ACSL4 and FTH1 in hippocampus. B-E Statistical graph of protein levels of GPX4, SLC7A11/ xCT, ACSL4 and FTH1. n = 5 per group. F Content of ferrous iron in the P24 hippocampus. n = 3 per group. G GSH concentration and the ratio of GSH and GSSH in the P24 hippocampus measured. n = 3 per group. H The MDA level in the P24 hippocampus. I The SOD concentration in the P24 hippocampus. J Representative immunofluorescence images of SLC7A11 (red) and DAPI (blue) in hippocampus. Scale bar = 100 μm. Mean fluorescence intensity of SLC7A11, n = 3 per group. K Representative immunofluorescence images of ACSL4 (red) and DAPI (blue) in hippocampus. Scale bar = 100 μm. Mean fluorescence intensity of ACSL4, n = 3 per group. L Representative immunofluorescence images of ROS (red) and DAPI (blue) in hippocampus. Scale bar = 100 μm. Mean fluorescence intensity of ACSL4, n = 3 per group. M Electron microscope of mitochondrial morphology in the P24 hippocampus. The second row represents a partial enlargement of the first row, respectively. Scale bar = 2 μm (raw) / 500 nm (zoom). Values was presented as mean ± SEM and analyzed by one-way ANOVA. ns > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The membranes were then incubated overnight at 4 °C with the following primary antibodies: GPX4 (1:2000; Cat# A11243; Abclonal), FTH1 (1:5000; Cat# A19544; Abclonal), ACSL4 (1:1000; Cat#38493T; Cell Signaling Technology), xCT/SLC7A11 (1:1000; Cat# 28001T; Cell Signaling Technology), and β-actin (1:50000; Cat# 66009-1-Ig; Proteintech).

Techniques: Concentration Assay, Immunofluorescence, Fluorescence, Microscopy

Journal: Biology of Sex Differences

Article Title: Sex differences in ferroptosis-related vulnerability to autism-like deficits in the adolescent medial prefrontal cortex following embryonic valproic acid exposure

doi: 10.1186/s13293-026-00982-x

Figure Lengend Snippet: E12.5 VPA induces ferroptosis-related alterations in the striatum without sex differences. A Representative protein bands of GPX4, SLC7A11/xCT, ACSL4 and FTH1 in striatum. B-E Statistical graph of protein levels of GPX4, SLC7A11/xCT, ACSL4 and FTH1. n = 5 per group. F Content of ferrous iron in the P24 striatum. n = 3 per group. G GSH concentration and the ratio of GSH and GSSH in the P24 striatum measured. n = 3 per group. H The MDA level in the P24 striatum. I The SOD concentration in the P24 striatum. J Representative protein bands of GPX4, SLC7A11/xCT and FTH1 in mPFC, hippocampus and striatum. K-M Statistical graph of protein levels of GPX4, SLC7A11/xCT and FTH1. n = 5 per group. N Representative immunofluorescence images of SLC7A11/xCT (red) and DAPI (blue) in striatum. Scale bar = 100 μm. Mean fluorescence intensity of SLC7A11/xCT, n = 3 per group. O Representative immunofluorescence images of ACSL4 (red) and DAPI (blue) in striatum. Scale bar = 100 μm. Mean fluorescence intensity of ACSL4, n = 3 per group. P Representative immunofluorescence images of ROS (red) and DAPI (blue) in striatum. Scale bar = 100 μm. Mean fluorescence intensity of ROS, n = 3 per group. Q Electron microscope of mitochondrial morphology in the P24 striatum. Scale bar = 2 μm (raw) / 500 nm (zoom). Values was presented as mean ± SEM and analyzed by one-way ANOVA. ns > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The membranes were then incubated overnight at 4 °C with the following primary antibodies: GPX4 (1:2000; Cat# A11243; Abclonal), FTH1 (1:5000; Cat# A19544; Abclonal), ACSL4 (1:1000; Cat#38493T; Cell Signaling Technology), xCT/SLC7A11 (1:1000; Cat# 28001T; Cell Signaling Technology), and β-actin (1:50000; Cat# 66009-1-Ig; Proteintech).

Techniques: Concentration Assay, Immunofluorescence, Fluorescence, Microscopy

Journal: Biology of Sex Differences

Article Title: Sex differences in ferroptosis-related vulnerability to autism-like deficits in the adolescent medial prefrontal cortex following embryonic valproic acid exposure

doi: 10.1186/s13293-026-00982-x

Figure Lengend Snippet: Comparable Ferroptosis-Associated alteration in the mPFC Following E12.5 and PND14 VPA Exposure. A Representative protein bands of GPX4, SLC7A11/xCT, ACSL4 and FTH1 in mPFC. B-E Statistical graph of protein levels of GPX4, SLC7A11/ xCT, ACSL4 and FTH1. n = 5 per group. F Content of ferrous iron in the P24 mPFC. n = 3 per group. G GSH concentration and the ratio of GSH and GSSH in the P24 mPFC measured. n = 3 per group. H The MDA level in the P24 mPFC. n = 3 per group. I The SOD concentration in the P24 mPFC. n = 3 per group. J Observation of the morphology of brain nerve cells in the mPFC by HE staining. Scale bar = 100 μm. The second row shows Nissl staining of neuronal morphology in the mPFC by. Scale bar = 100 (raw)/20 (zoom) µm. The black arrow represents cytoplasmic vacuolization, nuclear pyknosis and poorly defined nucleoli. Statistical graph of nissl body in the mPFC. n = 3 per group. K Representative immunofluorescence images of SLC7A11 (red) and DAPI (blue) in mPFC. Scale bar = 100 μm. Mean fluorescence intensity of SLC7A11, n = 3 per group. L Representative immunofluorescence images of ACSL4 (red) and DAPI (blue) in mPFC. Scale bar = 100 μm. Mean fluorescence intensity of ACSL4, n = 3 per group. M Representative immunofluorescence images of ROS (red) and DAPI (blue) in mPFC. Scale bar = 100 μm. Mean fluorescence intensity of ROS, n = 3 per group. N Electron microscope of mitochondrial morphology in the P24 mPFC. The second row represents a partial enlargement of the first row, respectively. Values was presented as mean ± SEM and analyzed by one-way ANOVA. ns > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The membranes were then incubated overnight at 4 °C with the following primary antibodies: GPX4 (1:2000; Cat# A11243; Abclonal), FTH1 (1:5000; Cat# A19544; Abclonal), ACSL4 (1:1000; Cat#38493T; Cell Signaling Technology), xCT/SLC7A11 (1:1000; Cat# 28001T; Cell Signaling Technology), and β-actin (1:50000; Cat# 66009-1-Ig; Proteintech).

Techniques: Concentration Assay, H&E Stain, Staining, Immunofluorescence, Fluorescence, Microscopy