Fig 1: LINC00641 ameliorated ketamine-induced neurological injury by activating the TrkB/PI3K/Akt signaling pathway. (A) Western blot was used to detect the expression variable of p-TrkB, p-PI3K and p-Akt protein and the expression of relative protein after miR-497-5pmimics was added to LINC00641 cell line. (B) Protein expression and relative protein expression levels of apoptosis-related proteins p-GSK-3β, p-CERB, Bcl-2, and Bax in miR-497-5p mimics were added to the overexpressing LINC00641 cell line. (C) The LINC00641-overexpressed cell line was treated with TrkB inhibitor (K-252a, 1.7 nM), and Western blot was used to detect p-TrkB, p-PI3K, and p-Akt expression of relative protein. *P < 0.05, **P < 0.01, and ***P < 0.001 vs. NC mimics group. #P < 0.05, ##P < 0.01, and ###P < 0.001 vs. miR-497-5p mimics + pcDNA3.1 group.
Fig 2: Enhanced phosphorylation of PI3K and Akt was reduced after cells with knockdown of TrKB expression. For both (A) HeLa and (B) SiHa cells, the phosphorylation of PI3K was obviously increased in cells from the model group compared to that in cells from the control group, whereas it was significantly decreased in cells from the siTrKB group compared with that in cells from the model group. However, no significant changes of PI3K protein were observed in the groups. For both (C) HeLa and (D) SiHa cells, the phosphorylation of Akt was clearly increased in cells from the model group compared to that in cells from the control group, whereas it was evidently decreased in cells from the siTrKB group compared with that in cells from the model group. However, no significant changes of Akt protein were observed in the groups. *P<0.05, vs. control; #P<0.05, vs. siTrKB.
Fig 3: Knockdown of CHRF reduces the protein expression level of PI3K/Akt pathway in LAD. Western blotting revealed that knockdown of CHRF decreased the protein expression levels of p-PI3K and p-Akt in (A) SPCA-1 and (B) NCI-H441 cells. Error bars represented the mean ± standard deviation of ≥3 independent experiments. **P<0.01 vs. control group. CHRF, cardiac hypertrophy-related factor; PI3K, posphoinositide-3-kinase; LAD, lung adenocarcinoma; p-, phosphorylated; T-total; si, small interfering RNA; NC, negative control.
Fig 4: Effect of overexpression of LINC00641 on neuronal injury in rats. (A–C) The expression of miR-497-5p and BDNF mRNA in overexpressing LINC00641 rats was detected by qPCR. (D–F) TUNEL and apoptosis index, ROS level, caspase-3 activity analysis. (G,H) Western blot was used to detect the protein expression and relative protein expression of p-PI3K and BDNF. KET, ketamine. *P < 0.05, **P < 0.01, and ***P < 0.001 vs. control group. #P < 0.05, ##P < 0.01, and ###P < 0.001 vs. KET + vector group. ∧P < 0.05, ∧∧P < 0.01, and ∧∧∧P < 0.001 vs. KET + LINC00641 group.
Fig 5: The study of the mechanism of CA on PI3K/AKT in the cellular induced by sevoflurane. The protein expression of p-PI3K, PI3K, p-AKT, and AKT was detected by WB. *p < 0.005, **p < 0.001, ***p < 0.0001.
Supplier Page from Abcam for Recombinant human PI3K (p110 delta/p85 alpha) protein (Active)