Figure 10. Inducible knock-down of hyLMN does not compromise the growth of Hydra. (A) Two shRNA constructs used for inducible knock-down of hyLMN - indHyLMN-hp405-929 and indHyLMN-hp1209-1707. (B) Transgenic polyps (indHyLMN-KD) and corresponding empty controls were incubated in 10 μM doxycycline (Dox) for 48 h prior to evaluation of the hyLMN knock-down efficiency by qRT-PCR. (C) hyLMN mRNA expression levels in two indHyLMN-KD hairpin405-929 lines (D1: n=4, and D10: n=3, mean±S.D.) and two indHyLMN-KD hairpin1209-1707 lines (F5: n=6, and F10: n=6) and in the respective controls. Asterisks indicate significant changes in expression (Mann-Whitney test); P values: D1 line = 0.039, D10 line = 0.022, F5 line < 0.001, F10 line = 0.004. (D) Growth curves for the indHyLMN-hp1209-1707 line F5 and control polyps (n=6 replicates, each five polyps on day 0; mean±S.D., linear regression lines with 95% CI corridors and goodness of fit R2). (E) Population doubling time values (mean±95% CI values) for the indHyLMN-hp1209-1707 line F5 and control polyps derived from the plot on D.