Fig 2: VEGFA is implicated in the miR-203a-3p-mediated PI3K/AKT signaling inhibition in HemECs. (A) Detection of PI3K and AKT phosphorylation by western blot analysis after miR-203-3p and VEGFA co-transfection. (B) Semi-quantification of p-PI3K/PI3K and p-AKT/AKT ratios among the different groups after miR-203-3p and VEGFA co-transfection. *P<0.05 and **P<0.01. VEGF, vascular endothelial growth factor; miR-203a-3p, microRNA-203a-3p; p-, phosphorylated; HemECs, human hemangioma endothelial cells.

Fig 3: A working model of wild-type IDH1 in primary GBM cells. IDH1 is the enzyme that catalyzes a-KG production from isocitrate. Wild-type IDH1 promotes the activation of the PI3K/AKT/mTOR pathway by affecting a-KG levels. IDH1 and a-KG may regulate primary GBM cell migration by modulating the PI3K/AKT/mTOR pathway. IDH1, isocitrate dehydrogenase 1; GBM, glioblastoma; a-KG, a-ketoglutarate; PI3K, phosphoinositide 3-kinase; AKT, protein kinase B; mTOR, mammalian target of rapamycin; P, phosphorylated.

Fig 4: Effect of FUS, alone and in combination with CDDO-me and/or nelfinavir, on both total and activated NF-?B and Akt protein levels in DU145 cells. The effect of combined treatment with nelfinavir and CDDO-me (CD), alone or in combination with FUS (N + CD + H4), on both total and activated (phosphorylated) NF-?B p65 (A–C) and Akt (D–F) protein levels is shown. Representative immunoblot images are in the top panel, along with the loading control (GAPDH). Bar graphs in each of the bottom panels of A–F show normalized densitometric values. Data are representative of three independent experiments (n = 3). Significant changes from untreated controls or individual treatment groups are shown as p-values (* p < 0.05; ** p < 0.01; and *** p < 0.001). Unlike the recurrence of NF-?B and Akt levels in DU145 cells exposed to FUS alone (cf. Supplemental Figure S1), our three-treatment combination (N + CD + H4) abrogated the reactivation of these two crucial transcription factors.

Fig 5: AKIP1 mediates NF-?B regulation via the Akt/IKKß pathway during the EMT. A, Serum-starved HeLa cells were pretreated with PI3K inhibitor Ly294002 (40 µM) or IKK inhibitor Bay117082 (15 µM) for 4 hours, followed by stimulation with AKIP1 protein (80 ng/mL) for 4 hours. Cell lysates were harvested and the levels of indicated proteins were measured using western blot analysis. B, pGL4.32 (luc2NF-?B-RE/Hygro) vector and pRL4-TK Renilla luciferase constructs were used for co-transfection of HeLa cells. The cells were further transfected 24 hours later with empty vector (EV) or AKIP1 expression plasmid. Some of the AKIP1-transfected cells were pretreated with Ly294002 (40 µM) or Bay117082 (15 µM) for 4 hours. The dual luciferase reporter assay was performed 24 hours following transfection of AKIP1. EV-transfected cells treated with TNFa (40 ng/mL) for 1 hour after transfection were used as the positive control. ** indicates P < .01. C, The HeLa cells were transfected with AKIP1 siRNA to knock down AKIP1. After 72 hours transfection, AKIP1 protein (80 ng/mL) was used to stimulate HeLa cells for 4 hours and the cell lysates were analysed using western blot