Extended Data Fig. 8: Expression and imaging controls for plant-related experiments.
a) Expression analysis of TWD40-1-β-GFP in independent plant lines in Col-0 background that were used for imaging. The stain free gel that serves as loading control is shown on top. The anti-GFP Western blot shows that the constructs are stably expressed. The Western Blot was done once. b) Bleed through control images for the data shown in Fig. 3d. The left and middle panels show the signal of a Col-0 root using our imaging conditions at different LUT levels as well as the FM4-64 channel image. Our imaging conditions, using a combination of three excitation lines for GFP does not result in FM4-64 bleed through. The scale bars equal 25 μm. This analysis was done for six independent plants. c) Expression analysis of independent TML-GFP and TML-AP-2μ-GFP lines in tml-1 background. The Western blot indicates both constructs are expressed at comparable levels. The Western Blot was done once. d) To control for random associations of TML-GFP (top panels) or TML-AP-2μ-GFP (bottom panels) with TPLATE-mSCARLET in the colocalization analysis, the green channel was rotated by 90° to the right and the images were re-analyzed. The pie chart quantifies the colocalization observed. Gray = colocalization, magenta = TPLATE only and green is TML only. Scale bars equal 5 μm. Two independent lines per construct were used and three plants were analysed per line. Of each root, six cells were used for the quantification.
