Prolonged incubation of Arabidopsis seedlings on microscopy glass upregulates autophagy in the epidermal root cells. (a) Arabidopsis seedlings expressing fluorescent marker for autophagy (pHusion-ATG8) in the wild-type (WT) or autophagy-deficient (atg5/7 KO) backgrounds were grown on standard Petri plates or in the RoPod v23. 5 days-old seedlings grown on a Petri plate were mounted between a standard microscopy-grade sample slide and a cover slip in a liquid 0.5xMS medium and incubated on the bench for the designated amount of time prior to imaging using confocal microscope. Seedlings of the same genotypes grown in the RoPod were imaged using the same settings (left panel). (b) Quantification of pHusion-positive puncta in the root cells illustrated in (a) reveals gradual upregulation of autophagic activity in the roots mounted on microscopy glass. The chart comprises representative data from one out of three individual experiments. Two-tailed t-test with unequal variants, n = 35; *, p-value < 0.05.