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Figure S1. Representative conversion in Kat8 after 520d-5p transfection. Figure S2. Comparative NGS analysis between hiPSC, 520d/hMSC progenitor cells, or hMSC. Figure S3. Comparative NGS analysis between hiPSC, 520d/hMSC progenitor cells, or hMSC regarding representative DNA repair genes. Figure S4. PCR array using RT2 Profiler PCR array system regarding epigenetics-related genes in 520d/HLF (5D, 7D) and hiPSC. (PDF 835 kb)

Summary of nucleotides alterations in respective genes regarding of our interest (Tables S1â S7). (DOCX 26 kb)

Subcutaneous changes of 520d-5p-expressing HMV-I in a xenograft model. Subcutaneous changes of 520d-5p-expressing HMV-I in a xenograft model were recorded as criteria for tumor formation and metastasis. In HMV-I, the disappearance rate of tumors was greater than the outcome (12.5Â %) of therapeutic administration in vivo. (PDF 32 kb)

A representative gene expression profile of 520d/HMV-I. Upregulation of p53 and Nanog and downregulation of AICDA were observed in 520d/HMV-I cells in vitro, similar to the results for 520d/HLF cells. *, Pâ

The suppression rate (%) of tumor volume. The suppression rate (%) of tumor volume compared with the control each week is shown in Table 1. Greater than 85 % suppression 6 weeks after administration was observed in a xenograft model by subcutaneous injection. (PDF 92 kb)

The side effects of 520d/atelocollagen and the presence of tumor cells in the inoculated tumor in seven organs. Neither the presence of embolism induced by atelocollagen or evidence of micrometastases was detected pathologically in brain, liver, spleen, heart, lung, small intestine and kidney. (PDF 185 kb)

Representative gene expression in HMV-I tumors without suppression. When the animals were sacrificed, most tumors grew increasingly and the suppressive effect of 520d-5p on tumor growth could not be observed by RT-PCR. *: Pâ

Effective rate of tumor disappearance. Effective rate (%) of tumor disappearance by systemic administration compared with that of the control is shown in Table 1. Greater than 75 % of the tumor cells disappeared based on macroscopic and microscopic observation. (PDF 94 kb)

The sequences of primers used in this study. Primers used in this study are presented. (PDF 69 kb)

A representative case 24Â weeks after subcutaneous injection. A pathologist recognized adipose tissue and skeletal muscle thought to be the existing structure but could not confirm the clear tumor composition at the injection site (HE stain; top left, the perspective of the specimen; bottom, x40, x100 magnification). We did not identify any macroscopic tumors in this case. (PDF 341 kb)

An in vivo study using glioblastoma cells (T98G) treated with miR-520d-5p (520d/T98G). Three surviving mice were examined for human-derived gene expression in murine brain tissue 3 months after intracranial injection. a KSN/Slc mice were anesthetized with sodium pentobarbital (50 mg/kg intraperitoneally) and placed in a stereotaxic apparatus. During surgery, the animals’ body temperature was maintained at 37 °C using a heating pad. The skull was exposed, and a small craniotomy was made over the left striatum. A 30-gauge injection needle connected to a 10-μl Hamilton syringe through polyethylene tubing was used for 520d/T98G cell transplantation. The injection needle was inserted stereotaxically into the left striatum (A 2.0 mm, L 0.5 mm, D 1.2 mm from bregma) (left), and 1 μl of cell suspension (1 × 108 cells/μl) was pressure-injected (right). After injection, the needle was slowly withdrawn, and the skull hole was covered with dental cement. The incision was sutured with 6-0 Prolene. After recovery from surgery, the animals were returned to their home cage. b Immunohistochemistry was performed using anti-hGFAP antibody, and human-derived GFAP protein expression was confirmed in glial cells and vascular endothelial cells in the murine thalamus (left to right: x40, x200, x400 magnification). (PDF 203 kb)