Fig. 1

NOTCH1 and EGFR signaling pathways were activated in human CRTC1-MAML2 fusion-positive MEC cells. a Western blot analysis was carried out on human CRTC1-MAML2-positive MEC cell lines (H3118, UM-HMC-3A, UM-HMC-3B, and H292) and a fusion-negative cells line (UM-HPA-1) using various antibodies as indicated. b Human MEC cells (H3118, UM-HMC-3B, and H292) were transduced with lentiviruses expressing shRNAs (shM2-1, shM2-3) that target the respective 3′ UTR and TAD domain of the CRTC1-MAML2 fusion and MAML2, or control shRNA (Ctl). Cells were collected at 96 h after viral infection and western blotting was performed. c Stable Cas9-expressing H3118 cells were transduced with lentiviruses expressing sgRNAs that target the exon 1 of MAML2 (sgMAML2-A, sgMAML2-B) or control (sgCtl). Cells were harvested at 96 h after viral infection and western blotting was performed. d Quantitative RT-PCR analysis of HES1 transcript levels in transduced MEC H3118 cells as described in (b) and (c) (n = 3; ns, p > 0.05; *p < 0.05; **p < 0.01). e Quantitative RT-PCR analysis of the transcript levels of three MAML co-activator family gene members, MAML1, MAML2, and MAML3 in human MEC H3118 cells