Phospho-Trk B (Tyr706) Antibody - #AF3461

Fig. 6. PLCβ4 is activated by mGluR1 to suppress the PIP2/PLCγ1/CREB/BDNF signaling pathway in hippocampal neurons. A) Immunoblots and quantification of mGluR1, PLCβ4, p-CREB and BDNF in neurons after 2 μM CdCl2 (Cd2) exposure, n = 3. B-E) The expression of mGluR1 in the hippocampus of male offspring following maternal Cd exposure at PND 21 B), PND 35 C), PND 56 D), and in old age E) was assessed by western blotting, n = 4/6/7/8. F-J) After transfection with 60 nM si-PLCβ4 for 24 h, a final concentration of 10 μM mGluR1 agonist (Ro-01–6128) was administered to hippocampal neurons for 24 h. F-J) Immunoblots and quantitative analysis of PLCβ4 G), p-PLCγ1 H), p-CREB (Ser133) I), and BDNF J), n = 4. K) After knocking down PLCβ4, hippocampal neurons were treated with an mGluR1 agonist for 24 h, and the mRNA levels of BDNF-I, BDNF-IV, and BDNF-VI were assessed by qPCR, n = 3. L) The representative images of Tuj1+ hippocampal neurons (red) treated with Ro-01–6128 before the knock-down of PLCβ4 (Scale bar: 50 µm) and the quantification of average neurite length, n = 3. After transfection with 60 nM si-PLCβ4 for 24 h, mGluR1 agonist (Ro-01–6128, 10 μM) and CdCl2 (2 μM) were administered to hippocampal neurons for 24 h. M-Q) The representative immunoblots M) and quantification of PLCβ4 N), p-PLCγ1 O), p-CREB (Ser133) P), and BDNF Q), n = 3. R-S) The representative images of MAP2+ hippocampal neurons (Scale bar: 50 µm) and the quantification of average neurite length, n = 3. The hippocampal neurons were transfected with si-PLCβ4 for 24 h, followed by the treatment of TrkB inhibitor ANA-12 (10 nM) with or without CdCl2 (2 μM) for 24 h. T, W) The expression levels of PLCβ4 and p-TrKB (Tyr706) were detected by Western blotting, n = 3/4. U-V, X-Y) The representative images of MAP2+ hippocampal neurons (Scale bar: 50 µm) and the quantification of average neurite length, n = 3. Data are represented as the mean ± SEM. Statistical significance are calculated by one-way ANOVA with Tukey’s post-hoc test and Student’s t test. * P 

Figure 3.| Effect of 7,8-DHF treatment on gene and protein levels of ERα and ERβ in both skeletal muscle and adipose tissues.(E)–(F) Western blotting analysis of skeletal muscle and adipose tissues isolated from mice that have been treated with 7,8-DHF for 24 weeks. Protein levels of ERα and its two phosphorylated forms p-S118 and p-Y537, the phosphorylation of TrkB and two key energy metabolism-related proteins, AKT and UCP1 were measured. Representative immunoblots and quantification are shown (n = 3 per group. *p <0.05, **p < 0.01, N.S: not significant, LFD control vs. LFD+DHF10, Student’s t test; &p < 0.05, &&p < 0.01, N.S: not significant, HFD control vs. HFD+DHF5, Student’s t test).