Knockdown of both CALCOCO1 and ZC3H10 rescues increased TMZ sensitivity in HOTAIR regulatory element KO glioma cells. (A) RT-qPCR verification of CALCOCO1 siRNA knockdown efficiency. Data represent means ± S.E.M. of three independent experiments. (**) P < 0.01 compared with Ctrl. (B) Western blot verification of CALCOCO1 siRNA knockdown efficiency. Relative protein levels were quantified by using ImageJ. (C) RT-qPCR verification of ZC3H10 siRNA knockdown efficiency. Data represent means ± S.E.M. of three independent experiments. (**) P < 0.01 compared with Ctrl. (D) Western blot verification of ZC3H10 siRNA knockdown efficiency. Relative protein levels were quantified by using ImageJ. (E) RT-qPCR verification of CALCOCO1 siRNA and ZC3H10 siRNA knockdown efficiency. Data represent means ± S.E.M. of three independent experiments. (***) P < 0.001, (****) P < 0.0001, compared with Ctrl. (F) Western blot verification of CALCOCO1 siRNA and ZC3H10 siRNA knockdown efficiency. Relative protein levels were quantified by using ImageJ. (G,H) LDH release was tested in control U251 glioma cells and HOTAIR regulatory element KO glioma cells transfected with control siRNAs and CALCOCO1 or ZC3H10 siRNAs. LDH release was tested after 48 h of TMZ (1 mM) treatment. Data represent means ± S.E.M. of three independent experiments. (I,J) Caspase 3/7 activities were tested in control U251 glioma cells and HOTAIR regulatory element KO glioma cells transfected with control siRNAs or CALCOCO1 or ZC3H10 siRNAs. Caspase 3/7 activities were tested after cells were treated 72 h with TMZ (1 mM). Data represent means ± S.E.M. of three independent experiments. (K) LDH release was tested in control U251 glioma cells and HOTAIR regulatory element KO glioma cells transfected with control siRNAs or CALCOCO1 and ZC3H10 siRNAs. LDH release was tested after cells were treated 48 h with TMZ (1 mM). Data represent means ± S.E.M. of three independent experiments. (*) P < 0.05 compared with corresponding Ctrl. (L) Caspase 3/7 activities were tested in control U251 glioma cells and HOTAIR regulatory element KO glioma cells transfected with control siRNAs or CALCOCO1 and ZC3H10 siRNAs. Caspase 3/7 activities were tested after cells were treated 72 h with TMZ (1 mM). Data represent means ± S.E.M. of three independent experiments. (*) P < 0.05 compared with corresponding Ctrl.
