Knockdown of Tcf7l1 induces differentiation of NE-4C cells toward neuron lineage cells in vitro. (A) Schematic showing the siRNA knockdown approach on the NE-4C cell line. (B) Cy3 signal indicating successful transfection of cells using siRNA. (C) qPCR results of Tcf7l1 in each group. Expression levels were normalized to Gapdh; three technical duplicates were performed in individual qPCR experiments. The t-test was used to determine statistical significance: (*) P ≤ 0.05, (**) P ≤ 0.01, (***) P ≤ 0.001, (****) P ≤ 0.0001, (ns) not significant. (D) qPCR assay of marker genes of different NSCs, progenitors, and neuroblasts under different treatment. Expression levels were normalized to Gapdh; three technical duplicates were performed in individual qPCR experiments. The t-test was used to determine statistical significance: (*) P ≤ 0.05, (**) P ≤ 0.01, (***) P ≤ 0.001, (****) P ≤ 0.0001, (ns) not significant. (E,F) Immunofluorescence staining against THBS4 (E) and DCX (F) in cultured cells under different treatments. Sections are counterstained with DAPI.
