phosphorylated p protein kinase b  (Proteintech)

Journal: Oncology Reports

Article Title: EIF3B-METTL3 complex promotes cell proliferation, invasion and EGFR/AKT signaling in cervical cancer

doi: 10.3892/or.2025.8936

Figure Lengend Snippet: EIF3B interacts with METTL3 and activates EGFR/AKT signaling in cervical cancer cells. (A) Representative western blots of co-immunoprecipitation assay. (B) Effect of EIF3B on EGFR mRNA expression in HeLa and SiHa cell lines. (C and D) Effects of EIF3B on EGFR protein and p-AKT expression. (C) Representative western blots and (D) quantification of EGFR protein and p-AKT expression in HeLa and SiHa cell lines. EIF3B, eukaryotic translation initiation factor 3B; METTL3, methyltransferase-like 3; p-, phosphorylated; si-, small interfering; oe-, overexpressing; NC, negative control. *P<0.05, **P<0.01 and ***P<0.001.

Article Snippet: The primary antibodies including EIF3B (1:5,000; cat. no. 10319-1-AP), METTL3 (1:10,000; cat. no. 15073-1-AP), N-cadherin (1:5,000; cat. no. 22018-1-AP), E-cadherin (1:5,000; cat. no. 20874-1-AP), Vimentin (1:20,000; cat. no. 10366-1-AP), EGFR (1:10,000; cat. no. 18986-1-AP), protein kinase B (AKT; 1:10,000; cat. no. 10176-2-AP), phosphorylated (p)-AKT (1:10,000; cat. no. 28731-1-AP) and GAPDH (1:10,000; cat. no. 10494-1-AP; all from Proteintech Group, Inc.) were incubated with the membrane overnight at 4°C.

Techniques: Western Blot, Co-Immunoprecipitation Assay, Expressing, Negative Control

Journal: Oncology Reports

Article Title: EIF3B-METTL3 complex promotes cell proliferation, invasion and EGFR/AKT signaling in cervical cancer

doi: 10.3892/or.2025.8936

Figure Lengend Snippet: EIF3B-METTL3 complex promotes EGFR/AKT signaling in cervical cancer cells. (A) Effect of the EIF3B-METTL3 complex on EGFR mRNA expression in HeLa and SiHa cell lines. (B and C) Effect of EIF3B-METTL3 complex on EGFR protein and p-AKT expression. (B) Representative western blots and (C) quantification of EGFR protein and p-AKT expression in HeLa and SiHa cell lines. EIF3B, eukaryotic translation initiation factor 3B; METTL3, methyltransferase-like 3; p-, phosphorylated; si-, small interfering; oe-, overexpressing; NC, negative control; ns, not significant. *P<0.05, **P<0.01 and ***P<0.001.

Article Snippet: The primary antibodies including EIF3B (1:5,000; cat. no. 10319-1-AP), METTL3 (1:10,000; cat. no. 15073-1-AP), N-cadherin (1:5,000; cat. no. 22018-1-AP), E-cadherin (1:5,000; cat. no. 20874-1-AP), Vimentin (1:20,000; cat. no. 10366-1-AP), EGFR (1:10,000; cat. no. 18986-1-AP), protein kinase B (AKT; 1:10,000; cat. no. 10176-2-AP), phosphorylated (p)-AKT (1:10,000; cat. no. 28731-1-AP) and GAPDH (1:10,000; cat. no. 10494-1-AP; all from Proteintech Group, Inc.) were incubated with the membrane overnight at 4°C.

Techniques: Expressing, Western Blot, Negative Control

Journal: Frontiers in Pharmacology

Article Title: The novel TFEB agonist desloratadine ameliorates hepatic steatosis by activating the autophagy-lysosome pathway

doi: 10.3389/fphar.2024.1449178

Figure Lengend Snippet: Desloratadine enhances the level of p-AMPK. (A,B) After treatment with 10 μM desloratadine (DLT) for 12 h or 24 h, HepG2 and L02 cells were subjected to Western blot analysis using the antibodies against p-Akt and p-p70S6K. β-actin was used as loading control. (C,D) The levels of p-AMPK were detected in the same desloratadine treated cells, and quantified by densitometric analysis and normalized to β-actin. (E) Assessment of TFEB-GFP nuclear localization. HeLa cells expressing TFEB-GFP were treated with Compound C (5 μM) and desloratadine (DLT) (10 μM) for 24 h and observed using a fluorescence microscope. Nuclei were stained with Hoechst 33,342 (blue). Scale bar, 20 μm. The percentage of cells with nuclear TFEB-GFP was quantified. (F,G) Effects of AMPK Inhibition on DLT-Induced nuclear translocation of TFEB. HepG2 and L02 cells were subjected to individual treatments with Compound C, Desloratadine (DLT), or a combination of both. Western blot was used to detect TFEB protein levels in nuclear and cytosolic fractions. GAPDH and histone H3 were used as loading controls. Densitometric analysis of Western blots was performed by ImageJ software. The protein level of TFEB was normalized to the level of cytoplasmic marker GAPDH or nuclear marker histone H3. (H,I) Western blot was used to measure the LC3-II levels after DLT treatment in presence of Compound C. β-actin was used as loading control. Data are presented as mean ± SD of three independent experiments (* P < 0.05, ** P < 0.01, ns, not significant).

Article Snippet: Antibodies against phosphorylated Akt (p-Akt) (1:500, sc-514032) and β-actin (1:500, sc-47778) were procured from Santa Cruz Biotechnology (Santa Cruz, California, United States).

Techniques: Western Blot, Control, Expressing, Fluorescence, Microscopy, Staining, Inhibition, Translocation Assay, Software, Marker

Journal: Frontiers in Pharmacology

Article Title: The novel TFEB agonist desloratadine ameliorates hepatic steatosis by activating the autophagy-lysosome pathway

doi: 10.3389/fphar.2024.1449178

Figure Lengend Snippet: The autophagy-lysosome pathway is activated in the liver of mice treated with desloratadine. (A) The protein levels of LC3, p-AMPK, p-Akt and p-p70S6K in the liver tissue were assessed by Western blot. Blots were quantified and normalized to β-actin (n = 5). (B) The mRNA levels of LC3B, p62, LAMP2, CTSB, CTSD and LIPA in liver tissue were detected by qRT-PCR analysis (n = 5). Data are presented as mean ± SD of three independent experiments (* P < 0.05, ** P < 0.01).

Article Snippet: Antibodies against phosphorylated Akt (p-Akt) (1:500, sc-514032) and β-actin (1:500, sc-47778) were procured from Santa Cruz Biotechnology (Santa Cruz, California, United States).

Techniques: Western Blot, Quantitative RT-PCR