Additional file 1 of MMP-3 cleavage of Lamin A induces pro-migratory nuclear deformity, nucleophagy, and their autophagic secretion with extracellular vesicles in metastatic cancer

Supplementary Material 1: Figure S1. Uncropped raw images of TEM of LuM1 tumoroid EVs and MMP3-KO EVs. Figure S2. Gene expression levels of MMP3 and LMNA positively correlated in clinical tumor specimens of HNSC and pancreatic adenocarcinoma. Figure S3. Original WB images of MMP-3, cleaved Lamin, and CD63. Figure S4. ImmunoEM images of Lamin A/C, cleaved Lamin, and negative control in EV particle fractions of LuM1 cells cultured in serum-free RPMI medium. Figure S5. ImmunoEM images of EpCAM, Lamin A/C, and cleaved Lamin in the EV particle fraction of LuM1 cells cultured in Essential 8 / RPMI medium. Figure S6. Amino acid sequences of Lamin A fragments and dimers that were bound with MMP-3 were identified by LC-MS/MS. Figure S7. CBB staining showing a new band at 30 kDa appearing upon MMP-3 overexpression and cellular heat shock stress at 43°C for 5 min. Figure S8. Original WB images of FL and truncated MMP-3. Figure S9. Original WB images of FL and cleaved Lamins, Cat, Act, and GAPDH. Figure S10. Quantification of Lamin A speckles and nuclear size changes. Figure S11. Original CLSM images of Lamin A/C and MMP-3 in starved LuM1pGFP cells. Figure S12. Original CLSM images of Lamin A/C and MMP-3 in starved M3pGFP cells. Figure S13. An original ICC image of cleaved Lamin (red), MMP3-GFP (green), and nuclei (blue) stained with DAPI. Figure S14. Original CLSM images of cleaved Lamin and MMP-3 in starved LuM1pGFP cells. Figure S15. Original CLSM images of cleaved Lamin and MMP-3 in starved M3pGFP cells. Figure S16. Original CLSM images of CD63 and LC3B in unstarved LuM1 and starved LuM1pGFP cells. Figure S17. Original CLSM images of CD63 and LC3B in starved M3pGFP cells. Figure S18. Original CLSM images of LC3B and MMP-3 in unstarved LuM1 and starved LuM1pGFP cells. Figure S19. Original CLSM images of LC3B and MMP-3 in starved M3pGFP cells. Figure S20. Original CLSM images of CD63 and MMP-3 in unstarved LuM1 and starved LuM1pGFP cells. Figure S21. Original CLSM images of CD63 and MMP-3 in starved M3pGFP cells. Figure S22. Original CLSM images of cleaved Lamin and CD63 in unstarved LuM1 and starved LuM1pGFP cells. Figure S23. Original CLSM images of cleaved Lamin and LC3B in starved LuM1pGFP cells. Figure S24. Original CLSM images of MMP-3 and Lamin A/C in Colon26, LuM1, and MMP3-KO cells. Figure S25. Original CLSM images used for quantification of MMP3-dependent lamina loss and nuclear atypia. Figure S26. Representative full-focused ICC images of Lamin A/C and MMP-3 in Colon26, LuM1, and MMP3-KO cells. Figure S27. Representative 3D images of MMP-3 and Lamin A/C in Colon26, LuM1, and MMP3-KO cells. Figure S28. Representative auto full-focus ICC images of a tumor-like cell aggregate (tumoroid) of LuM1 cells. Figure S29. Floating bar graphs showing Lamin A/C expression level (A) and area (B) per cell, quantified from ICC images. Figure S30. Kaplan-Meier analysis of MMP3-high (red) and low (black) expression patients with HNSC, PDAC, rectum and lung ACs. Figure S31. Kaplan-Meier analysis of LMNA-high (red) and low (black) expression patients with HNSC, PDAC, rectum and lung ACs. Table S1. Rates of MMP-3 (+) nuclei and cleaved Lamin (+) nuclei in patient-derived tumor tissues of stage I HNSC. Table S2. Rates of MMP-3 (+) nuclei and cleaved Lamin (+) nuclei in patient-derived tumor tissues of stage IV HNSC