Fig. 6

ThermomiR-377-3p improved the sensitivity of NPC cells and cancer stem-like cells to hyperthermia in vitro by directly suppressing Cirbp expression. A-C qRT-PCR assay for detecting the expression of selected miRNAs of which Cirbp might be a potential target gene in the indicated NPC cells treated with or without hyperthermia at 40 °C and 42 °C for 30 min. ThermomiRs (i.e., miR-143 and miR-142-5p) were used as positive controls. D, E qRT-PCR assay for detecting the expression of miR-377-3p D and Cirbp E in NPC cells transiently transfected with miR-377-3p mimics or inhibitor. F Western blot was employed to detect Cirbp expression in NPC cells transiently transfected with miR-377-3p mimics or inhibitor. G Diagram of 3’-UTR-WT and 3’-UTR-MUT of Cirbp containing reporter constructs. H Luciferase reporter assays in HEK293T cells co-transfected with WT or MUT 3’-UTR and miRNAs as indicated. I, J Colony formation assay I and tumor sphere formation assay J were performed in miR-377-expressing NPC cells treated with or without hyperthermia at 42 °C for 30 min. K EdU assay was performed in NPC cells transiently transfected with miR-377-3p mimics and then treated with or without hyperthermia at 42 °C for 30 min. L Western blot was employed to detect Cirbp expression in miR-377- and Cirbp-expressing NPC cells. M, N Colony formation assay M and tumor sphere formation assay N were performed in miR-377- and Cirbp-expressing NPC cells treated with or without hyperthermia at 42 °C for 30 min