Fig 1: Effects of DHT on the expression of PSD95 and the phosphorylation of ERK1/2, eIF4E in HT22 cells after ZIP9 overexpression. (A, B) The overexpression efficiency of ZIP9 protein in HT22 cells infected with ZIP9-oe or nc-oe. (C–F) Representative Western blot (C) and quantification (D–F) of PSD95 (D), and the phosphorylation of ERK1/2 (E) and eIF4E (F) in HT22 cells. (G, J) Representative IF staining images (J) and quantification (G) of PSD95 in HT22 cells. (H, K) Representative IF staining images (K) and quantification (H) of p-ERK1/2 in HT22 cells. (I, L) Representative IF staining images (L) and quantification (I) of p-eIF4E in HT22 cells. Scale bars = 20 µm. DHT, dihydrotestosterone; ERK1/2, Extracellular signal-regulated kinase ½; eIF4E, Eukaryotic translation initiation factor 4E; ZIP9, Zrt-, Irt-like protein 9; PSD95, postsynaptic density protein 95. (*P < 0.05, n = 5).
Fig 2: ZIP9 mediated the effects of DHT on the phosphorylation of eIF4E and the expression of PSD95 in HT22 cells pretreated with eFT508. (A-C) Representative Western blot (A) and quantification (B, C) of the Phosphorylation of eIF4E (B) and expression of PSD95 (C) in HT22 cells pretreated with eFT508. (D, F) Representative IF staining images (F) and quantification (D) of p-eIF4E in HT22 cells pretreated with eFT508. (E, G) Representative IF staining images (G) and quantification (E) of PSD95 in HT22 cells pretreated with eFT508. Scale bars = 20 µm. DHT, dihydrotestosterone; ZIP9, Zrt-, Irt-like protein 9; PSD95, postsynaptic density protein 95, ERK1/2, Extracellular signal-regulated kinase ½; eIF4E, Eukaryotic translation initiation factor 4E; eFT, eFT508. (*P < 0.05, n = 5).
Fig 3: Effects of DHT on the phosphorylation of ERK1/2 and eIF4E in Tfm male mice and HT22 cells after ZIP9 knockdown. (A–C) Representative Western blot (A) and quantification (B, C) of the phosphorylation of ERK1/2 (B) and eIF4E (C) in the hippocampus of the four groups of mice. (D, E) The knockdown efficiency of ZIP9 protein in HT22 cells infected with ZIP9-shRNA or nc-RNA. (F-I) Representative Western blot (F) and quantification (G-I) of the expression of PSD95 (G), and the phosphorylation of ERK1/2 (H), eIF4E (I) in HT22 cells. (J, K) Representative IF staining images (K) and quantification (J) of PSD95 in HT22 cells. (L, M) Representative IF staining images (L) and quantification (M) of p-ERK1/2 in HT22 cells. (N, O) Representative IF staining images (N) and quantification (O) of p-eIF4E in HT22 cells after ZIP9 knockdown. Scale bars = 20 µm. IF, immunofluorescence; DHT, dihydrotestosterone; Tfm, Testicular feminization mutation; ERK1/2, Extracellular signal-regulated kinase ½; eIF4E, HT22, hippocampal neuron cells; ZIP9, Zrt-, Irt-like protein 9; PSD95, postsynaptic density protein 95. (*P < 0.05, n = 5).
Fig 4: ZIP9 mediated the effects of DHT on the phosphorylation of ERK1/2, eIF4E and Expression of PSD95 after ZIP9 overexpression. (A–D) Representative Western blot (A) and quantification (B–D) of the phosphorylation of ERK1/2 (B), eIF4E (C) and the expression of PSD95 (D) in HT22 cells pretreated with SCH772984. (E, H) Representative IF staining images (H) and quantification (E) of p-ERK1/2 in HT22 cells. (F, I) Representative IF staining images (I) and quantification (F) of p-eIF4E in HT22 cells. (G, J) Representative IF staining images (J) and quantification (G) of PSD95 in HT22 cells. Scale bars = 20 µm. DHT, dihydrotestosterone; ZIP9, Zrt-, Irt-like protein 9; PSD95, postsynaptic density protein 95, ERK1/2, Extracellular signal-regulated kinase ½; eIF4E, Eukaryotic translation initiation factor 4E. S, SCH772984. (*P < 0.05, n = 5).
Fig 5: ZIP9 mediated the effects of DHT on the phosphorylation of ERK1/2 and eIF4E in APP/PS1 mice hippocampus. (A, B) Representative Western blot (A) and quantification (B) of the phosphorylation of ERK1/2 in the hippocampus of the four groups of mice. (C, D) Representative Western blot (C) and quantification (D) of the phosphorylation of eIF4E in the hippocampus of the four groups of mice. DHT, dihydrotestosterone; ZIP9, Zrt-, Irt-like protein 9; ERK1/2, Extracellular signal-regulated kinase ½; eIF4E, Eukaryotic translation initiation factor 4E. (*P < 0.05, n = 6).
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