rabbit anti phosphorylated p38 mapk  (Cell Signaling Technology Inc)

Figure Legend Snippet: Fig. 4. Detection of iron content, ROS levels and activation of PI3K-Akt/MAPK signaling pathway in the hippocampus of mice. (A-D) Immunofluorescence staining was used to detect Fpn1+tdTomato+ cells (A-B), FtL+tdTomato+ cells (C) and FtH+tdTomato+ cells (D) in the cKO-Td and Cre-Td mice. Scale bar = 50 μm. Yellow arrows indicate merged cells. Dot lines indicate the SGZ region. (E-H) Western blot was used to detect the expression levels of FtL and FtH (E-F), and Bcl2 and Bax (G-H) in cKO and Control mice. Data are shown as mean ± SEM. n = 5. (I) ROS levels in hippocampus were detected by ROS assay kit (n = 10). (J) The mRNA levels of PI3K and Akt were detected by qRT-PCR (n = 5). (K-O) Western blot was used to detect the expression of p-PI3K, p-Akt, p-ERK and p-p38 MAPK (n = 5). Data are shown as mean ± SEM. p values were determined by two-tailed Student’s t tests. *p < 0.05; * *p < 0.01; * **p < 0.001.

Techniques Used: Activation Assay, Immunofluorescence, Staining, Western Blot, Expressing, Control, ROS Assay, Quantitative RT-PCR, Two Tailed Test

Figure Legend Snippet: Fig. 5. Effects of FAC supplementation on the proliferation of NSCs and PI3K-Akt/MAPK signaling pathways in the hippocampus of wild-type mice. (A) Flow diagram of experimental procedures. (B-G) Immunofluorescence or immunohistochemical staining was used to detect Ki67+ cells (B-C), BrdU+ cells (D-E) and Nestin+ signals (F-G) in SGZ and quantitative analysis (n = 5). Scale bar = 50 μm. (H-L) Western blot was used to detect the expression of p-PI3K, p-Akt, p-p38 MAPK and p-ERK (n = 5). Data are shown as mean ± SEM. p values were determined by two-tailed Student’s t tests. *p < 0.05; * *p < 0.01; * **p < 0.001.

Techniques Used: Protein-Protein interactions, Immunofluorescence, Immunohistochemical staining, Staining, Western Blot, Expressing, Two Tailed Test

Figure Legend Snippet: Fig. 6. Effects of FAC or H2O2 treatment on the proliferation of cultured NSCs and PI3K-Akt/MAPK signaling pathways. (A-B) Representative images and diameter analysis of the neurospheres of primary hippocampal NSCs treated with different concentrations of FAC (A) or H2O2 (B). n = 50–103 neurospheres per group. (C) CCK-8 kit was used to detect the effect of Fpn1 siRNA treatment on the proliferation of NE-4C stem cells. n = 6. (D) Western blot was used to detect the activation of p-Akt in NE-4C cells treated with SiFpn1-2. n = 3. (E-F) CCK-8 kit was used to detect the effect of FAC (E) or H2O2 (F) on the proliferation of NE-4C stem cells. n = 9. (G-J) Western blot was used to detect the expression of p-Akt in NE-4C stem cells treated with different concentrations of FAC (G-H) or H2O2 (I-J). n = 3. (K) CCK-8 kit was used to detect the effect of ROS inhibitor NAC on the proliferation of FAC-treated NE-4C stem cells (n = 11). (L-N) Western blot was used to detect the expression of FtH and p-Akt in NE-4C cells treated with ROS inhibitor with or without FAC supplementation (n = 3). (O) CCK-8 kit was used to detect the effect of PI3K inhibitor (LY294002) and MAPK inhibitor (U0126) on the proliferation of FAC-treated NE-4C cells (n = 8). Data are shown as mean ± SEM. p values were determined by one-way ANOVA tests in Fig. A-C, E, F, H and J. p values were determined by two-way ANOVA in Fig. K, M-O. *p < 0.05; * *p < 0.01; * **p < 0.001; ns, non-significant. Each sample in the experiments had two to four replicates, and three independent experiments were performed.

Techniques Used: Cell Culture, Protein-Protein interactions, CCK-8 Assay, Western Blot, Activation Assay, Expressing