Fig 2: Gene expression analysis of transfected hUC-MSCs incubated in oligo induction medium: qPCR analysis showing the transcriptional activation of OLIG2 and MYT1L genes in (A) OLIG2, (B) MYT1L, and (C) synergistic (OLIG2 + MYT1L) transfected groups. Lineage specification analysis indicated higher expression level of GFAP and OLIG2 compared to NES, demonstrating the glial fate specification. Transcriptional analysis of oligodendrocyte-specific genes, i.e., SOX10, NKX2.2, GALC, CNP, CSPG4, MBP, and PLP1, presenting their significant overexpression compared to the control group after 3 days of incubation in oligo induction medium. Each experiment was run in triplicates and acquired data were analyzed through an independent sample t-test at statistical significance level; p = 0.05 (p < 0.05 = *, p < 0.01 = **, and p < 0.001 = ***). NS = not significant.

Fig 3: Gene expression analysis of transfected hUC-MSCs incubated in normal medium: qPCR analysis indicating the transcriptional activation of OLIG2 and MYT1L genes in (A) OLIG2, (B) MYT1L, and (C) synergistic (OLIG2 + MYT1L) transfected groups. Lineage specification analysis presented higher expression levels of GFAP and OLIG2 in comparison to NES, demonstrating the glial fate specification. Transcriptional analysis of oligodendrocyte-specific genes, i.e., SOX10, NKX2.2, GALC, CNP, CSPG4, MBP, and PLP1, presenting their significant overexpression compared to the non-transfected control group after 3 days of incubation in normal medium. Each experiment was run in triplicates and acquired data were analyzed through an independent sample t-test at statistical significance level; p ≤ 0.05 (p < 0.05 = *, p < 0.01 = **, and p < 0.001 = ***). NS = not significant.

Fig 4: Protein expression/immunocytochemical analysis of transfected hUC-MSCs in normal and oligo induction media after 7 days: (A) Immunocytochemical analysis of OLIG2, MYT1L, and synergistic (OLIG2 + MYT1L) transfected groups, indicating positive expression of MYT1L, OLIG2, NG2, and MBP proteins. (B,C) Fluorescent intensities showing intense expression of oligodendrocyte-specific proteins, i.e., MYT1L, OLIG2, NG2, and MBP in all the transfected groups as compared to the non-transfected control group after 7 days of incubation in both normal and oligo induction media. However, the OLIG2-transfected group exhibited reduced fluorescence signals of MBP in the normal medium. The fluorescent values (n = 30) were analyzed through an independent sample t-test at statistical significance level; p = 0.05 (p < 0.05 = *, p < 0.01 = **, and p < 0.001 = ***). NS = not significant.