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. 2021 Dec 9;12:7150. doi: 10.1038/s41467-021-27384-4

Fig. 5. Extracellular Otk is cleaved in EEs by Mmp1 after tissue injury.

Fig. 5

A Full-length Otk with a GFP tag on the C-terminal, intracellular region was overexpressed specifically in EEs to visualize intracellular Otk. Antibodies against the N-terminal, extracellular Otk was used to visualize extracellular Otk. Both N-terminal and C-terminal regions of Otk were detected on the EE cortex in undamaged tissue (white arrowhead), but only C-terminal Otk was detected after tissue damage (yellow arrowhead). B N-terminal Otk was detected in EEs from undamaged tissue (white arrowhead), but was largely absent in EEs from damaged tissue (yellow arrowhead). Depleting Mmp1 in EEs from damaged tissue prevented the loss of N-terminal Otk (white arrowhead). N-terminal Otk was detected in ISCs from both undamaged and damaged tissue (white arrows). mean ± SD; n = sample size as follows, A (Relative N-term Otk intensity): 30 cells from 6 flies from 2 independent experiments, A (%Otk-GFP + EEs): 6 flies from 2 independent experiments, B: 7 flies from 2 independent experiments; N.S. = not significant (A = 0.4043, B = 0.7252), ***P < 0.001 (A, B < 0.0001), based on two-tailed Student’s t test (A, B) and one-way ANOVA with Dunnett test (B). Scale bar = 10 µm. Source data are provided as a Source Data file.