Fig. 5: Latent membrane protein 1 (LMP-1) and nicotine suppressed vacuolar protein sorting 33B (VPS33B) expression through phosphoinositide-3 kinase (PI3K)/AKT/c-Jun signaling. | Cell Death & Disease

Fig. 5: Latent membrane protein 1 (LMP-1) and nicotine suppressed vacuolar protein sorting 33B (VPS33B) expression through phosphoinositide-3 kinase (PI3K)/AKT/c-Jun signaling.

From: VPS33B interacts with NESG1 to modulate EGFR/PI3K/AKT/c-Myc/P53/miR-133a-3p signaling and induce 5-fluorouracil sensitivity in nasopharyngeal carcinoma

Fig. 5

a Quantitative polymerase chain reaction (qPCR) assay indicated that VPS33B was downregulated in the VPS33B-overexpressing HONE1 and SUNE1 cells after transduction of LMP-1 plasmids. Student’s t test, mean ± SD, *P < 0.05, **P < 0.01. b, c 3-[4,5-Dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (b) and 5-ethynyl-2′-deoxyuridine incorporation (c) assays were performed to demonstrate the impact of miR-133a-3p on the proliferation of A549 and H1975 cells. Student’s t test, one-way analysis of variance, mean ± SD, *P < 0.05, **P < 0.01. Scale bar: 100 μm. d Changes in PI3K/AKT/p-PI3K/p-AKT and VPS33B expression were detected by western blot analysis in VPS33B-overexpressed HONE1 and SUNE1 cells after transduction of LMP-1 plasmids. β-Actin was used as a loading control. e, f The mRNA level of VPS33B were downregulated in the nicotine-treated HONE1 and SUNE1 cells in different concentrations (0.1, 1, 10, 100 μmol/L) at 72 h or at different times (24, 48, 72, 100, 132, and 132 h) in 10 μmol/L via qPCR assay. g Changes in PI3K/AKT/p-PI3K/p-AKT, c-Jun, and VPS33B expression were detected by western blot analysis in nicotine-treated A549 and H1975 cells after transfection of PI3K inhibitor Ly294002. β-Actin was used as a loading control. h VPS33B was indicated to be upregulated in nicotine-treated HONE1 and SUNE1 cells after transfection of PI3K inhibitor Ly294002 by qPCR assay. Student’s t test, mean ± SD, *P < 0.05, **P < 0.01. i The binding of c-Jun with VPS33B promoter was examined in nicotine-treated HONE1 and SUNE1 cells after transfecting Ly94002 by qPCR assay. Student’s t test, mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001

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