Figure 6. Extracellular Islr attenuates epithelial Hippo signaling to activate YAP signaling in vitro .

1. Schematics of culture epithelial cells with supernatant from HEK293FT cells transfected with pcDNA3.1 empty vector or pcDNA3.1‐hISLR‐HA plasmids. Western blotting for HA in the supernatant from HEK293FT cells transfected with the indicated plasmids.
2. Gene set enrichment for YAP/TAZ/TEAD direct target genes in transcriptome profiles for HEK293FT cells treated by hISLR‐HA‐enriched supernatant.
3. Heatmap showing the altered Hippo‐related genes in the transcriptome profiles. The parameter of the color key indicated the fold changes converted to log2 of gene fragments per kilobase per million mapped reads (FPKM) values.
4. Western blotting for pMST1/2, MST1, pMOB1 and MOB1, pYAP, and YAP in lysates from HEK293FT cells cultured in the supernatants with or without hISLR‐HA. β‐actin was used as a loading control.
5. Western blotting for HA and ISLR in the supernatant from NCM460 colonic epithelial cells with pcDNA3.1 empty vector or pcDNA3.1‐hISLR‐HA plasmids. Western blotting for pMST1/2, MST1, pMOB1 and MOB1, pYAP, and YAP in lysates from NCM460 CRC cells cultured in the supernatants with or without hISLR‐HA. β‐actin was used as a loading control.
6. Immunofluorescence for YAP in HEK293FT cells cultured in the supernatant with or without hISLR‐HA. Scale bar: 10 μm.
7. Western blotting for YAP in cytoplasmic or nuclear proteins from HEK293FT cells cultured in the supernatant with or without hISLR‐HA. Lamin a/c, a positive control for nuclear proteins. α‐Tubulin, a positive control for cytoplasmic proteins.
8. qRT–PCR for CTGF and CYR61 in HEK293FT cells cultured in the supernatant with or without hISLR‐HA; n = 3.

Data information: In (H), data are presented as mean ± SD. ***P < 0.001 (Student's t‐test).Source data are available online for this figure.