A Krüppel-like factor establishes cellular heterogeneity during schistosome tegumental maintenance
Fig 5
Knockdown of klf4 alters the heterogeneity of the sm13
+ TRL. (A) FISH result showing klf4 RNAi causes no significant effect on the number of cells expressing sm13 (left), quantification of the number of sm13+ cells per mm of worm (right). Control (RNAi) n= 28, klf4 (RNAi) n=26. (B) qPCR detection of the expression of sm13 (n=10 experiments) and up-regulated DEGs including ludp (Smp_319910) (n=10 experiments), ly6e (Smp_334920) (n=6 experiments) and litaf (Smp_333330) (n=10 experiments) following klf4 RNAi. (C) FISH results showing a significant increase in the number of ludp+ cells following klf4 RNAi (left); Double FISH (inset region indicated by the dotted rectangle from the left panel) showing expression of sm13 relative to the ludp+ cells (right); the Venn diagram in upper right shows the percentage of sm13+ cells expressing ludp. White arrows indicate the sm13+ludp+ cells, red arrows indicate the sm13-ludp+ cells. Scale bar, 100 µm. (D) Quantification of the number of ludp+ cells per mm of worm. Control (RNAi) n=34, klf4 (RNAi) n=38. (E) Quantification of percentage of sm13+ cells expressing ludp. Control (RNAi) n=20, klf4 (RNAi) n=21. (F) Double FISH showing expression of calpain relative to the ludp+ cells, and the Venn diagram in upper right shows the percentage of calpain+ cells expressing ludp. Control (RNAi) n=19, klf4 (RNAi) n=13. Scale bar, 10 µm. (G) Quantification of percentage of calpain+ cells expressing ludp. Scale bar, 10 µm. Data are presented as mean ± SD. Welch’s t-tests were performed in Panel A, D, E and G, and multiple paired t-tests were performed in Panel B.