Figure 3.

Smed-Tim is a gene of the planarians circadian clock. (A) Planarians were submitted to the condition L/D and then Smed-Tim expression was evaluated using RT-qPCR. Planarians have a maximum of Tim expression in dark phase. The results are presented as the mean ± SD (3 worms per time point processed individually in triplicate, number of experiments = 3). (B) Planarians were submitted to D/D cycle and Smed-Tim expression was evaluated using RT-qPCR. Worms submitted to D/D condition have a desynchronized Smed-Tim expression cycle. The results are presented as the mean ± SD (3 worms per time point processed individually in triplicate, number of experiments = 3). (C) The expression of Smed-Tim was evaluated at CT20 in worms submitted to L/D condition and infected by S. aureus. (D) Whole-mount in situ hybridization analysis of the expression of Smed-Tim in control and S. aureus infected animals were fixed at CT 20 (representative micrographs of 2 experiments; 5 animals per experimental conditions; scale bars, 250 µm). See also Figure S3. (E) Melatonin level was determined between CT14 and CT22 in animals eGFP (RNAi) and Smed-Tim (RNAi) under the L/D cycle level was assessed in planarian controls or silenced for Smed-Tim via RNAi. The results are presented as the mean ± SD (30 worms per time point, number of experiments = 2, *p < 0.05).