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. 2019 May 30;177(6):1405–1418.e17. doi: 10.1016/j.cell.2019.05.011

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© 2019 The Author(s)

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Figure S6 — The Influence of RCO and STM on Leaf Growth, Related to Figure 4

(A-B) Heat-maps of growth anisotropy for A. thaliana RCOg (A), and C. hirsuta rco (B) leaves (white lines indicate the orientation of cell growth where anisotropy is higher than 40%).

(C-D) Cell lineage tracing analysis in the leaf blade (left panels) or petiole/midrib (right panels) for A. thaliana RCOg (C) and C. hirsuta rco (D) leaves from 3-7 DAI. Colors show the correspondence between cells at 3 DAI and their clonal sectors at 7 DAI.

(E-F) Heat-maps of cell proliferation for A. thaliana RCOg (E) and C. hirsuta rco (F) leaves.

(G-H) Heat-maps of cell size for A. thaliana RCOg (G) and C. hirsuta rco (H) leaves.

(I-J) Heat-maps of growth anisotropy for A. thaliana BLS::STM (I) and C. hirsuta stm (J) leaves (white lines indicate the orientation of cell growth where anisotropy is higher than 40%). As the BLS::STM sample used for 3 DAI in (M) and (N) was not captured at 1 DAI; the heat-map of anisotropy over 1-3 DAI in (I) was obtained from an independent time-lapse series.

(K-L) Cell lineage tracing analysis in the leaf blade (left panel) or petiole/midrib (right panel) for A. thaliana BLS::STM (K) and C. hirsuta stm (L) leaves from 3-7 DAI. Colors show the correspondence between cells at 3 DAI and their clonal sectors at 7 DAI.

(M-N) Heat-maps of cell proliferation for A. thaliana BLS::STM (M) and C. hirsuta stm (N) leaves.

(O-P) Heat-maps of cell size for A. thaliana BLS::STM (O) and C. hirsuta stm (P) leaves.

(Q-R) Heat-maps of area extension in an A. thaliana BLS::STM transgenic leaf (Q) and a C. hirsuta stm mutant (R). Asterisk indicates additional protrusion emerging from the leaf margin in (Q).

(S-V) Quantifications of mean area extension, cell proliferation, cell size, and stomata density for A. thaliana RCOg (S), C. hirsuta rco (T), A. thaliana BLS::STM (U) and C. hirsuta stm (V) compared to WT leaves (error bars indicate SEM). (S) A. thaliana RCOg, n = 2-4 (stomata density) and n > 150 (remaining plots, 4 independent time-lapse series). (T) C. hirsuta rco, n = 3 (stomata density) and n > 80 (remaining plots, 3 independent time-lapse series). (U) A. thaliana BLS::STM, n = 3-4 (stomata density) and n > 100 (remaining plots, 4 independent time-lapse series). (V) C. hirsuta stm, n = 2-4 (stomata density) and n > 300 (remaining plots, 4 independent time-lapse series).

(W) pAtBLS::3xGFP expression pattern in A. thaliana (left column) and pChSTM::3xVENUS expression pattern in C. hirsuta (right column). Confocal images with GFP or VENUS signal in yellow and propidium iodide staining in magenta.

Note that tissue-dependent growth patterns are still observed in leaf primordia of A. thaliana RCOg (C) and BLS::STM (K) plants, as well as C. hirsuta rco (D) and stm (L) mutants.

DAI indicates days after primordia initiation. Scale bars, 100 μm in (A-B, E-J, M-R, and W bottom panels), 20 μm in (C-D, K-L, and W top panels).