Fig. 5

Knockdown of IMPDH2 in cells without cytoophidia affects proliferation rate. a Wild-type, mut-IMPDH2 Col_5 and mut-IMPDH2 Col_32 were transfected with Non-Target siRNA (NT) or 100 nM IMPDH2 targeting siRNA for 48 h and labelled with EdU and anti-IMPDH2 antibody. Proportion of EdU-positive cells is presented in the top of each panel. More than 1000 cells were counted and at least three independent experiments were performed for each group. b IMPDH1 and 2 protein levels in WT and mut-IMPDH2 Col_5 and Col_32 indicating a decrease in protein levels under increasing doses of siRNA for IMPDH2-Kd. c The relative IMPDH activity of cell lysates of wild-type and mutant cell lines. d Proliferation rate for each cell line in various conditions was normalized with Non-Target siRNA transfected groups. Cells were transfected with 100 nM IMPDH2 siRNA for IMPDH2-Kd and supplemented with a daily dose of 1 mM guanosine (Gua) or 1 mM GTP. Error bars means SD. Student’s t-test, ***P ≤ 0.001. e Intracellular level of GTP (standardized with ATP) of wild-type and mutant cells after transfection of Non-Target or IMPDH2 siRNA