The Integrator complex regulates differential snRNA processing and fate of adult stem cells in the highly regenerative planarian Schmidtea mediterranea
Fig 3
Expression of integrator genes is increased in neoblasts and is essential for UsnRNA processing.
qRT-PCR quantification of transcript levels. (A, B) Depletion of Integrator complex subunits results in a significant accumulation of 3’-unprocessed UsnRNAs in both intact animals and in the neoblast-enriched X1 FACS fraction. (C, D) ints3 and ints9 mRNA levels were significantly higher in the X1 fraction compared to both the X2 and the Xins fraction, and were strongly down-regulated in γ-irradiated animals depleted of neoblasts. (E) 3’-unprocessed U2A2 snRNA, relative to the total nascent U2A2 level, was less abundant in RNA of X1 cells compared to that of Xins cells (left chart; average of the X1/Xins ratios of 3 biological replicates; p = 0.02, two-sided heteroscedastic t-test) and accumulated substantially in X1 cells upon ints RNAi (right chart). Expression levels are the averages of three biological replicates normalized to those of the total pool of the respective UsnRNA variant in A+B, to those of gapdh in C+D and to those of total nascent U2A2 in E (error bars represent standard deviation; two-sided t-test, * p<0.05, ** p<0.01, *** p<0.001).