Skip to main content
. 2022 Aug 4;11:e80778. doi: 10.7554/eLife.80778

Figure 5. Expression of E2F and its targets during oral apparatus (OA) regeneration.

(A) E2F is an early-expressed gene. Plot shows expression of E2F homolog SteCoe_12750 as a function of time including in cycloheximide-treated cells. OA (blue curve) indicates expression in sucrose-shocked cells. The purple curve shows that E2F is still upregulated in cells in which protein synthesis is inhibited. (B) Expression heatmap of predicted E2F targets in the OA-specific expression module. Clusters refer to clusters in the original clustering of OA-specific genes (Figure 2A). Cluster 1 is not included in the plot because there were no predicted E2F targets among the cluster 1 genes. (C–F) Examples of control cells at 4, 6, 8, and 24 hr after regeneration. Purple arrows mark the location of oral primordium. Red arrow head marks the location of a fully formed OA. (G–J) Examples of E2F RNAi cells at 4, 6, 8, and 24 hr, illustrating the proper formation of the oral primordium by 4 hr, but lack of visible primordium or OA at later stages. All scale bars 200 μm. (K) Fraction of cells regenerating properly at each time point. For the 4, 6, and 8 hr time points, on-schedule regeneration is defined as cells having a visible oral primordium (4 hr), a full length primordium extended along the side of the cell (6 hr), and an oral apparatus located at the anterior end of the cell whose diameter is at least 50% of the diameter of the cell (8 and 24 hr). Total number of cells analyzed from five separate biological experiments was n=206 for control and 310 for RNAi. Error bars represent standard error for the proportions expected based on a Bernoulli distribution. For all four time points, the proportion of RNAi cells regenerating on schedule (defined according to a set of milestone events as described in Materials and methods) was significantly less than for controls, with p values of (p=0.007, p<0.0001, p<0.0001, and p=0.0005) respectively based on Fisher’s exact test.

Figure 5.

Figure 5—figure supplement 1. Additional information for Figure 5.

Figure 5—figure supplement 1.

Outcomes of regeneration assays for E2F RNAi experiments shown for five separate experiments done on different days with different batches of cells prepared separately. Numbers of cells analyzed in each experiment are as follows. (A) Control n=49 RNAi n=24. (B) Control n=49 RNAi n=64. (C) Control n=47 RNAi n=115. (D) Control n=43 RNAi n=71. (E) Control n=18 RNAi n=36. Data from these experiments were combined to produce the overall histogram in Figure 5K. Error bars represent standard error.