Fig 3. Evaluation of microfilament configurations and stomatal apertures over a diurnal cycle.

(A) Changes in stomatal aperture at 2-hour increments over a 24 h (12 h light/12 h dark) diurnal cycle. (B) Changes in population distributions of each class of actin MF configuration obtained by hierarchical cluster analysis over a 12 h light/12 h dark cycle. Changes in actin MF configuration over a 12 h light/12 h dark diurnal cycle in metrics for (C) orientation, (D) bundling, and (E) density of MFs. (F) Representative images of stomatal guard cells collected by spinning disc confocal microscopy during light and dark periods over a 24 h diurnal cycle. Images were collected at 5 h Zeitgeber time (lights on, stomata maximally opened) and 19 h Zeitgeber time (lights off, stomata maximally closed), respectively. The averaged values of apertures at each time point are shown inside images. Error bars represent means ± SEM from stomatal aperture and metrics for MF orientation, bundling and density. Data was collected from 3 independent biological repeats (n = 42–52).