Estrogen Receptor-beta Antibody - #AF6469

Figure 7. Genistin interacts with ERβ to regulate megakaryocyte differentiation. (A) ERα or ERβ and ligands (genistin) by molecular docking. (B) Lysates from Meg-01 cells were incubated with or without genistin (200 μM) for 2 h, and different concentrations of pronase E (1:500, 1:1000, or 1500) were added for 10 min at 40°C, ERβ content was detected using western blot analysis. (C) Lysates from Meg-01 cells were incubated with genistin (100, 200, 400 μM) for 2 h, and a final concentration of pronase E (1:1000) was added for 10 min at 40 °C. ERβ expression was detected by western blot analysis. (D) Lysates from Meg-01 cells were incubated with genistin (200 μM) for 9 h, then reacted at different temperatures (45, 48, 51, 54, 57 °C) for 5 min, and finally the ERβ content was detected by western blot. (E) Lysates from Meg-01 cells containing or without different doses of genistin (100, 200, 400 μM) were incubated for 9 h, and then reacted at 48 °C for 5 minutes, and finally the ERβ content was detected by western blot. (F-G) Meg-01 cells were detected by flow cytometry using genistin (20 μM), PHTPP (20 μM), and both, respectively. The histogram shows the percentage of CD41+CD42b+ cells for each group. (H-I) PI staining in Meg‑01 cells was used to analyze the DNA content (2, 4 and ≥ 8 N) by flow cytometry after genistin (20 μM), PHTPP (20 μM), and both interventions, respectively. All data represents the mean±SD of three independent experiments; *p < 0.05, **p < 0.01, ***p < 0.001, vs the Ctrl group; #p < 0.05, ##p < 0.01, ###p < 0.001, vs the Gen group; Gen: genistin, Ctrl: control.

Figure 8. Genistin can promote PI3K/AKT and MEK/ERK expression. (A-G) The expression levels of ERβ/GAPDH, HSP90/GAPDH, p-PI3K/PI3K, p-AKT/AKT, p-MEK/MEK, and p-ERK/ERK were detected by western blot after Meg-01 cells were treated with genistin for 5 days. (H) Representative immunofluorescence images of ERβ fluorescently double-stained with p-ERK and p-AKT, respectively, on bone marrow cells after 10 days of treatment with genistin in a mouse model of radiation thrombocytopenia. All data represents the mean ± SD of three independent experiments; *p < 0.05, **p < 0.01, ***p < 0.001, vs the Ctrl group; Gen: genistin, Ctrl: control.

Figure 3. Spatial colocalization of substance P with ERβ in the spinal cord was investigated using dual-label confocal immunofluorescence examination. The specificities of the immunopositive staining of ERβ (green) and substance P (red) were demonstrated. Substance P colocalized with ERβ in the dorsal horn of the spinal cord, mainly in the laminae I and II, a connection site of pain transmission. (A) Coronal sections through the spinal cord of DLBP mice. Scale bar: 200 μm. (B) The magnified dorsal horn areas of the spinal cord of DLBP mice. Scale bar: 20 μm.

Fig. 3. The expression of SGK1, ENaC-α, ERβ and PR in decidua tissue was detected by immunohistochemistry. Note: vs Control group, ***P < 0.001. Scale bars: a 100 μm, b 50 μm (n = 10).

Figure 6.Effect of sulpiride (SUL) on fasting-induced changes in protein expression. Figures represent the changes in the protein expression of RASD Family Member 2 (RASD2), brain-derived neurotrophic factor (BDNF), cAMP-response element binding protein (CREB), phospho-CREB (p-CREB), protein kinase B (Akt), dopamine D2 receptor (DRD2), estrogen receptor α (ERα), and estrogen receptor β (ERβ) in the hippocampus (HP) (A, a–h), prefrontal cortex (PFC) (B, i–p), and striatum (C, q–x) in ovariectomized mice. The data are expressed as mean ± SEM (n = 3–5). Two-way ANOVA with Tukey’s honestly significant difference (HSD), *P < .05 vs ovariectomy (OV), **P < .01 vs vehicle (VEH):non-fasting; #P < .05, ##P < .01 vs VEH:fasting.

Figure 5 The expression level of estrogen receptors (ERs) in femur and spine. (A) Western blotting results of expression level of ERα and ERβ in femoral and spine growth plate (GP), n = 3. (B–E) Semiquantitative analysis of ER expression levels. (F) Quantitative real-time PCR results of ER genes in femoral and vertebral GP, n = 4. (G–J) Immunohistochemistry (IHC) results of expression level of ERα and ERβ in femoral and vertebral GP, n = 6. Scale bar: 20 μm. (K) Comparison of ERα HSCORE between femoral and spine GP (FGP, femoral growth plate; SGP, spine growth plate). (L, M) Results of Spearman’s analysis between ERα HSCORE and femur/spine length. *p < 0.05, ***p < 0.001 and ****p < 0.0001; ns denotes not significant.

Figure 5 The expression level of estrogen receptors (ERs) in femur and spine. (A) Western blotting results of expression level of ERα and ERβ in femoral and spine growth plate (GP), n = 3. (B–E) Semiquantitative analysis of ER expression levels. (F) Quantitative real-time PCR results of ER genes in femoral and vertebral GP, n = 4. (G–J) Immunohistochemistry (IHC) results of expression level of ERα and ERβ in femoral and vertebral GP, n = 6. Scale bar: 20 μm. (K) Comparison of ERα HSCORE between femoral and spine GP (FGP, femoral growth plate; SGP, spine growth plate). (L, M) Results of Spearman’s analysis between ERα HSCORE and femur/spine length. *p < 0.05, ***p < 0.001 and ****p < 0.0001; ns denotes not significant.