Fig 2: Expression of trilineage differentiation markers. A) Matched iPSCs and differentiated endoderm, mesoderm, ectoderm cells were analyzed by immunostaining followed by flow cytometry for expression of markers specific for each germ layer (mean ± SEM, n = 7–10 cell lines). See also Fig. S1 for isotype control data, Fig. S3 for morphological changes after differentiation, Fig. S4 for paired data, and Fig. S5 for independent validation of iPSC NES staining. B) Representative flow cytometry plots showing co-immunostaining of pluripotency marker POU5F1 with each of the four candidate ectoderm markers in undifferentiated iPSCs. See also Fig. S6 for complete dataset. C, D) Expression of pluripotent marker POU5F1 (C) and four candidate ectoderm markers (D) quantified by qPCR in matched iPSCs and ectoderm differentiated cells (mean ± SEM, n = 8). Gene expression is shown as fold change of ectoderm differentiated cells compared to iPSCs, with dotted line indicating fold change of 1. * P < .05, *** P < .001, **** P < .0001, two-tailed paired t-tests.

Fig 3: Validation of blood-derived human iPSCs. Live stain of representative iPSC colonies showing A) alkaline phosphatase activity, and B) SSEA-4 expression. C) Flow cytometry analysis of pluripotency markers POU5F1 and TRA-1–60, and differentiation marker SSEA-1 in iPSCs (mean ± SD, n = 30). See also Fig. S1 for flow cytometry data on isotype controls. D) Karyotype analysis of three iPSC lines showed normal chromosomal content. See also Fig. S2 for additional characterization of pluripotency marker expression and genomic aberrations.

Fig 4: miRNA was abundantly secreted into the culture supernatant. The expression level of (a) miR-302b, (b) LIN28, and (c) Oct4 in 105 iPSCs and 105 RPE cells and in 2 mL of their culture supernatants. For LIN28 and Oct4, the vertical axis is the relative expression level as the expression level in RPE cells was 1. The level of miRNAs in the supernatant was corrected by multiplying the detected value with 20, because only 100 µL in 2 mL of the culture supernatants was used as a sample. As the cells were extracted from whole cells, such a correction was not performed. UD: no miRNA detected; error bar = + 3SD.

Fig 5: M2 macrophage induced by IGF2BP3 enhence the stemness of glioblastoma stem cells (GSCs). (A, B) Representative images showing neurosphere formation GSC11/23 cocultured with THP1 (Mφ) transfected with plasmids overexpressing nonsense sequence or full‐length IGF2BP3 cDNA and transfected with miR‐NC or miR‐1246 mimics and si‐NC, si‐IGF2BP3#2 or si‐IGF2BP3#3 (scale bar, 200 μm). (C, D) Flow cytometry were performed to analyze the proportion of CD133+OCT4+GSC11 cocultured with macrophages transfected with plasmids overexpressing nonsense sequence or full‐length IGF2BP3 cDNA and transfected with miR‐NC or miR‐1246 mimics and si‐NC, si‐IGF2BP3#2 or si‐IGF2BP3#3. (E–J) Flow cytometry was applied to measure the surfaces marker (CD133) of GSC23 cocultured with THP1 (Mφ) transfected with plasmids overexpressing nonsense sequence or full‐length IGF2BP3 cDNA and transfected with miR‐NC or miR‐1246 mimics and si‐NC, si‐IGF2BP3#2 or si‐IGF2BP3#3, and quantification was performed. Data depict the mean ± standard deviation and are representative of three independent experiments (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001).