Fig 2: IR affects protein levels of the IKK members, upstream activators, and phosphorylation of the downstream p65 NFκB in A549 and H1299 lung cancer cell lines. A549 and H1299 cells were treated with 4 Gy IR, and cytoplasmic (IKKα, pIKKα, IKKγ, IκBα, pp65 NFκB, RAF1, and pAKT1S1) or nuclear fractions (pp65 NFκB) of cell lysates were collected 30 min, 4 and 24 h, and 7 days post-irradiation and analyzed with Western blot. Non-irradiated samples were used as an untreated control (denoted as 0), and β-actin and lamin B1 were used as loading controls for cytoplasmic and nuclear fractions, respectively.

Fig 3: miR-98 inhibits IGF1R/k-Ras/Raf/MEK/ERK signaling pathway. (A) SO-RB50 and Y79 cells were transfected with miR-98 mimics, mimics NC, pcDNA-vector or pcDNA-IGF1R and k-Ras, p-Raf/1, Raf/1, p-MEK1/2, MEK1/2, p-ERK1/2 and ERK1/2 were measured using western blot analysis. β-actin served as an internal control. (B and C) Immunohistochemistry was conducted to detect k-Ras, p-MEK1/2 and p-ERK1/2 expression in RB tissues with high or low miR-98 expression (magnification, ×200). Bar graphs demonstrated a significant inverse association between miR-98 and k-Ras, p-MEK1/2 and p-ERK1/2 expression in RB tissues (n=20). Data are presented as the mean ± standard deviation of three individual experiments. ERKK, extracellular signal-regulated kinase; IGF1R, insulin-like growth factor-1 receptor; MEK, mitogen activated protein kinase kinase; miR, microRNA; p, phosphorylated.

Fig 4: Activation of the MAPK pathway by coexpression of BPTF and Raf1. (A) Western blot analysis of MAPK pathway-related proteins in cancerous lymph nodes or adjacent normal tissues from TCL patients. (B) mRNA and protein expression levels of BPTF in Hut-102 cells following transfection with NC or shBPTF. (C) mRNA and protein expression levels of BPTF in Hut-102 cells transfected with NC (empty vector) or with a BPTF-overexpressing vector. (D) Western blot analysis of MAPK pathway-related in Hut-102 cells transfected with NC or shBPTF. (E) The interaction between BPTF and MAPK pathway-related proteins was analyzed using the STRING website. (F) Correlation analysis of BPTF and Raf1 expressions from TCGA database. (G) mRNA and protein expression levels of Raf1 in Hut-102 cells overexpressing BPTF. (H) mRNA and protein expression levels of Raf1 in Hut-102 cells following BPTF silencing. **P<0.01 and ***P<0.001. BPTF, bromodomain PHD finger transcription factor; TCL, T-cell lymphoma; NC, negative control; sh, short hairpin; MAP2K1, mitogen-activated protein kinase kinase 1; MAPK1, mitogen-activated protein kinase 1; GRB2, growth factor receptor bound protein 2; RASGRP1, RAS guanyl releasing protein 1.

Fig 5: miR-489 promotes apoptosis in glioma cells by attenuating RAF1-Bax-mediated cell survival pathways. (A and B) U87 and (C and D) U251 cells were transfected with control, miR-489 mimic or miR-489 inh for 24 h, and subsequently stained with Hoechst 33258 and observed under a fluorescence microscope. Flow cytometry analysis was performed to measure the apoptosis in transfected cells after 24 h. (E) U87 cells were transfected with control, miR-489 mimic or miR-489 inh and western blotting was used to determine the expression of the indicated proteins at 24 h post-transfection. U87 cells were transfected with control, (F) miR-489 mimic or (G) miR-489 inh and the mRNA expression levels of the indicated genes were measured at 24 h after transfection. (H) U251 cells were transfected with control, miR-489 mimic or miR-489 inh and western blotting was used to determine the expression of the indicated proteins at 24 h after transfection. U87 cells were transfected with control, (I) miR-489 mimic or (J) miR-489 inh, and the mRNA expression levels of the indicated genes were measured at 24 h after transfection. **P<0.01 vs. control. Data are presented as the mean ± standard deviation of three independent experiments. A two-way ANOVA was used to analyze the differences between two groups. miR-489 inh, miR-489 inhibitor; miR-489, miR-489 mimic; con, control.