Supplementary Figure 2: Screen of different auxin receptor F-box proteins using miniAID degron in human cells.
From: An efficient auxin-inducible degron system with low basal degradation in human cells

(a) Scheme for screening different auxin receptor F-box proteins using miniAID-mEGFP tagged seipin as the target. (b) Relative mCherry intensities analyzed by FACS in cells expressing different mCherry-tagged auxin receptor F-box proteins. Bars represent mean; n=2 independent experiments. At: Arabidopsis thaliana; Gh: Gossypium hirsutum; Mn: Morus notabilis; Nc: Noccaea caerulescens; Os: Oryza sativa. (c) Live cell Airyscan images showing subcellular localization of mCherry-tagged auxin receptor F-box proteins. This experiment was repeated once independently with similar results. (d) FACS plots showing overlays of GFP histograms at 0 h (blue line), 1 h (green line) and 16 h (red line) IAA in cells expressing the nontagged (left) or mCherry-tagged (right) constructs indicated. Figures exemplifying the gating strategy with SSC, FSC for cells, and trigger pulse width for singlets are shown above. Ctrl: no auxin receptor F-box protein, WT: wildtype A431 cells without GFP, blue dotted line and red dotted line: drawings for comparison of GFP peaks. This experiment was repeated once independently with similar results. (e) Mean GFP intensities in (d). Bars represent mean ± s.d.; n=4 biologically independent samples, except for ctrl (n=2). Individual data points of different auxin receptor F-box proteins are combinations of nontagged and mCherry-tagged constructs.