Journal: Antioxidants (Basel, Switzerland)

Article Title: Blue Light Alters the Composition of the Jejunal Microbiota and Promotes the Development of the Small Intestine by Reducing Oxidative Stress.

doi: 10.3390/antiox11020274

Figure Lengend Snippet: Figure 9. Effects of PI3K inhibitor, AKT inhibitor, GSK-3β inhibitor, GPR43 agonist, GPR41 agonist, Gi inhibitor and Gq inhibitor on p-PI3K protein expression (A), p-AKT/total-AKT ratio (B), p-GSK- 3β/total-GSK-3β ratio (C), β-catenin protein expression (D), cyclin D1 protein expression (E) and CIECs proliferation stimulation index (F), Bax protein expression (G), Bcl-2 protein expression (H) and Caspase-3 protein expression (I). LY294002 is an inhibitor of PI3K; GSK-690693 is an inhibitor of AKT; CHIR-99021 is an inhibitor of GSK-3β; 4-CMTB is an agonist of GSK-3β; AR420626 is an agonist of GPR41; PTX is an inhibitor of Gi and Ym254890 is an inhibitor of Gq. Different letters show significant differences among the various treatment groups (p < 0.05). p-PI3K: phosphorylated PI3-kinase; p-AKT: phosphorylated AKT; p-GSK-3β: phosphorylated GSK-3β. CIECs: chick small intestinal epithelial cells.

Article Snippet: Next, we used 5% skimmed milk to blocked nitrocellulose membranes for 1 h. Subsequently, the primary antibodies, including anti-Claudin-1 (rabbit, 1:1000; Invitrogen, Carlsbad, CA, USA), anti-Occludin (rabbit, 1:1000;, Invitrogen, Carlsbad, CA, USA), anti-ZO-1 (rabbit, 1:1000; Invitrogen, Carlsbad, CA, USA), anti-phospho-PI3 Kinase (Tyr458) antibody (rabbit, 1:1000, CST, Boston, MA, USA), anti-phosphor-AKT (Ser473) antibody (rabbit, 1:500, CST, Boston, MA, USA), anti-AKT antibody (rabbit, 1:500, CST, Boston, MA, USA), anti-phosphor-GSK-3β antibody (rabbit, 1:500, Abnova, Taiwan, China), anti-GSK-3β antibody (rabbit, 1: 500, Proteintech Group, Inc, Wuhan, China), anti-β-catenin antibody (rabbit, 1:1000, Proteintech Group, Inc, Wuhan, China), anti-cyclinD1 antibody (mouse, 1:200, Abbexa, Cambridge, UK), anti-Bax antibody (goat, 1:1000, Biorbyt, NJ, USA), anti-Bcl-2 antibody (rabbit, 1:1000, Biorbyt, NJ, USA), anti-Caspase-3 antibody (rabbit, 1:1000, CST, Boston, MA, USA) and anti-β-actin (mouse, 1:4000; Co Win Biotech Co., Inc, Beijing, China), were incubated with the membranes overnight at 4 ◦C.

Techniques: Expressing

Journal: International immunopharmacology

Article Title: S100A8 knockdown activates the PI3K/AKT signaling pathway to inhibit microglial autophagy and improve cognitive impairment mediated by chronic sleep deprivation.

doi: 10.1016/j.intimp.2024.113375

Figure Lengend Snippet: Fig. 6. S100A8 knockdown inhibits excessive autophagy in the hippocampus of CSD mice A: Autophagosomes in the hippocampus examined by transmission electron microscopy microscopy. The autophagosomes are denoted by arrows. Scale bar: 1 µm. B-E: An Immunohistochemical staining assay was performed to detect the LC3B, P62 and Beclin-1 levels in hippocampal tissues. Scale bars: 200 and 20 µm F: Detection of S100A8 and apoptosis-related proteins in the hippocampal tissues of mice in each group by Western blot analysis. G: Western blot analysis of the protein expression of phosphorylated PI3K, Akt and P70S6K in hippocampal tissue H- N: Analysis of the protein band gray intensity. n = 5–6 mice per group; *P < 0.05 vs Control; #P < 0.05 vs CSD; &P < 0.05 vs Si-NC + CSD; $P < 0.05 vs Si-S100A8 + CSD.

Article Snippet: Antibodies against GFAP (#3670), LC3B (#83506), phosphophosphoinositide 3-kinase (PI3K) p85 (#4228), phospho-Akt (#13038), PI3K p85 (#4292), Akt (#4685), phospho-P70S6K (#97596), and P70S6K (#9202) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Knockdown, Transmission Assay, Electron Microscopy, Microscopy, Immunohistochemical staining, Staining, Western Blot, Expressing, Control

Journal: International immunopharmacology

Article Title: S100A8 knockdown activates the PI3K/AKT signaling pathway to inhibit microglial autophagy and improve cognitive impairment mediated by chronic sleep deprivation.

doi: 10.1016/j.intimp.2024.113375

Figure Lengend Snippet: Fig. 7. S100A8 knockdown activates the PI3K/AKT signaling pathway to inhibit CSD-induced aberrant autophagy and apoptosis A: Representative HE-stained images of hippocampal tissue from each group of mice. Arrows indicate areas of neuronal loss and altered morphology. Scale bars: 100 and 20 µm. B-G: Immu- nofluorescence analysis of LC3B, P62 and beclin-1 in the hippocampal tissues of mice in each group. Scale bars: 200 and 20 µm. H: Determination of apoptosis and autophagy-related protein expression in the hippocampal tissues of mice after the addition of a PI3K/Akt inhibitor by WB. I-N:Analysis of the protein band - gray intensity. n = 5–6 mice per group; *P < 0.05 vs Control; #P < 0.05 vs CSD; &P < 0.05 vs Si-NC + CSD; $P < 0.05 vs Si-S100A8 + CSD.

Article Snippet: Antibodies against GFAP (#3670), LC3B (#83506), phosphophosphoinositide 3-kinase (PI3K) p85 (#4228), phospho-Akt (#13038), PI3K p85 (#4292), Akt (#4685), phospho-P70S6K (#97596), and P70S6K (#9202) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Knockdown, Staining, Expressing, Control

Journal: International immunopharmacology

Article Title: S100A8 knockdown activates the PI3K/AKT signaling pathway to inhibit microglial autophagy and improve cognitive impairment mediated by chronic sleep deprivation.

doi: 10.1016/j.intimp.2024.113375

Figure Lengend Snippet: Fig. 9. S100A8 knockdown activates the PI3K/AKT signaling pathway to inhibit rapamycin-induced aberrant autophagy and apoptosis A-F: Expressions levels of autophagy-related proteins LC3B, P62 and Beclin-1 in microglia were detected by immunofluorescence staining. Scale bars: 50um. G: Autophagy and apoptosis- related protein expression levels in microglia after addition of PI3K/Akt inhibitors were detected by Western blot. H-M: Analysis of the protein band gray inten- sity. n = 3/group. *P < 0.05 vs Control; #P < 0.05 vs RAPA; &P < 0.05 vs Si-NC + RAPA; $P < 0.05 vs Si-S100A8 + RAPA.

Article Snippet: Antibodies against GFAP (#3670), LC3B (#83506), phosphophosphoinositide 3-kinase (PI3K) p85 (#4228), phospho-Akt (#13038), PI3K p85 (#4292), Akt (#4685), phospho-P70S6K (#97596), and P70S6K (#9202) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Knockdown, Immunofluorescence, Staining, Expressing, Western Blot, Control

Journal: International immunopharmacology

Article Title: S100A8 knockdown activates the PI3K/AKT signaling pathway to inhibit microglial autophagy and improve cognitive impairment mediated by chronic sleep deprivation.

doi: 10.1016/j.intimp.2024.113375

Figure Lengend Snippet: Fig. 10. S100A8 aggravates abnormal autophagy and apoptosis in mouse hippocampus by inhibiting PI3K/AKT signaling.

Article Snippet: Antibodies against GFAP (#3670), LC3B (#83506), phosphophosphoinositide 3-kinase (PI3K) p85 (#4228), phospho-Akt (#13038), PI3K p85 (#4292), Akt (#4685), phospho-P70S6K (#97596), and P70S6K (#9202) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques:

Journal: Hereditas

Article Title: Corynoxine suppresses lung adenocarcinoma proliferation and metastasis via inhibiting PI3K/AKT pathway and suppressing Cyclooxygenase-2 expression.

doi: 10.1186/s41065-024-00343-x

Figure Lengend Snippet: Fig. 4 Corynoxine reduces COX-2 levels through PI3K/ATK pathway suppression. After treated with different concentrations of Corynoxine, (a) COX-2 mRNA expression was detected by qRT-PCR. (b) Western immunoblotting was used to detect COX-2 and β-actin levels. (c) After treated with different concentrations of Corynoxine or Celecoxib (40 µM) for 24 h, the level of PEG2 was evaluated by ELISA assay. (d) A549 cells were treated with indicated doses of Corynoxine for 24 h after pretreatment with the COX-2 selective inhibitor Celecoxib (40 µM) for 8 h, and the cell viability was determined by CCK-8 assay. After treated with different concentrations of Corynoxine. (e) Different subunits of PI3K (PIK3CA, PIK3CB and PIK3CD) were detected by qRT- PCR. (f) Western immunoblotting was used to detect PI3K p110δ, p-AKT, AKT, and β-actin levels. (g) After treatment with 100 µM Corynoxine or 10 µM LY294002 for 24 h, western immunoblotting was used to assess the expression of COX-2 and β-actin expression. Data were expressed as mean ± SD, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: Blots were incubated overnight with primary antibodies prepared in 5% BSA (Solarbio, SW3015, China) at 4 °C, including antibodies specific for Vimentin (1:1000, Abcam, ab20346, USA), Bcl-2 (1:2000, Proteintech, 26593-1-AP, China), Bax (1:200, Santa Cruz, sc-20067, USA), E-cadherin (1:1000, CST, 14472, USA), PI3K p110δ (1:200, Santa Cruz, sc-55589, USA), AKT (1:1000, CST, 4685, USA), p-AKT (1:1000, CST, 4060, USA), and COX-2 (1:1000, CST, 12282, USA), and β-actin (1:5000, TransGen, TC201, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, CCK-8 Assay

Journal: Hereditas

Article Title: Corynoxine suppresses lung adenocarcinoma proliferation and metastasis via inhibiting PI3K/AKT pathway and suppressing Cyclooxygenase-2 expression.

doi: 10.1186/s41065-024-00343-x

Figure Lengend Snippet: Fig. 6 Corynoxine suppresses intratumoral PI3K/AKT/COX-2 pathway activity. (a) Bax, Bcl-2, and β-actin protein levels. (b) E-cadherin, Vimentin, and β-actin protein levels. (c) Intratumoral COX-2, PI3K, AKT, p-AKT, and β-actin levels were detected. Data were expressed as mean ± SD, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: Blots were incubated overnight with primary antibodies prepared in 5% BSA (Solarbio, SW3015, China) at 4 °C, including antibodies specific for Vimentin (1:1000, Abcam, ab20346, USA), Bcl-2 (1:2000, Proteintech, 26593-1-AP, China), Bax (1:200, Santa Cruz, sc-20067, USA), E-cadherin (1:1000, CST, 14472, USA), PI3K p110δ (1:200, Santa Cruz, sc-55589, USA), AKT (1:1000, CST, 4685, USA), p-AKT (1:1000, CST, 4060, USA), and COX-2 (1:1000, CST, 12282, USA), and β-actin (1:5000, TransGen, TC201, China).

Techniques: Activity Assay