Fig. 12. Cells derived from a clone transformed with F-GFP plasmid were subjected to gene silencing of all the c-subunuits or the F-subunit itself (a-i); 16, 25 and 40 hours after the beginning of feeding, samples were taken and fixed. Control cells (a-c) show no difference in fluorescence within 40 hours, in contrast to cells after silencing of the c-subunits, which causes displacement of the F-subunit from the contractile vacuole complex after 16 hours (d). After 25 hours, only remnants of the contractile vacuole complex are visible and these have disappeared after 40 hours of feeding. Silencing of the F-subunit shows very rapid elimination of the fluorescence of F-GFP already after 16 hours of feeding (g). (j-l) Non-injected control cells showing autofluorescence typical for Paramecium. Silencing of the F-subunit as well as the c-subunits reduces fluorescence of F-GFP to the level of autofluorescence within 40 hours. Bar, 10 µm.