Lama2 And Samsn1 Mediate the Effects of Brn4 on Hippocampal Neural Stem Cell Proliferation and Differentiation

Figure 3

Lama2 activation or Samsn1 overexpression inhibits the proliferation of hippocampal NSCs. (a) Quantification of Lama2 mRNA level normalized to β-actin in the hippocampal NSCs which were transfected with different concentrations of Lama2 CRISPR SAM coactivated plasmid. (b) Quantification of Samsn1 mRNA level normalized to β-actin in the hippocampal NSCs which were infected by Samsn1 overexpression lentivirus. (c) The upper panel is the representative immunoblots of Samsn1 and β-actin after being infected by Samsn1 overexpression lentivirus; the lower panel is the quantification of Samsn1 protein level normalized to β-actin. (d) The upper panel is the representative immunoblots of Lama2 and β-actin after being infected by Lama2 CRISPR SAM coactivated plasmid; the lower panel is the quantification of Lama2 protein level normalized to β-actin. (e, f) Representative EdU-staining in primary hippocampal NSCs. . (g, h) Quantification of EdU-positive cells normalized to Hoechst. (i–k) Representative cell cycle distribution analyzed by flow cytometry in primary hippocampal NSCs after Lama2 activation (i, j) and quantification for NSCs in the different stages of the cell cycle (k). (l–n) Representative cell cycle distribution analyzed by flow cytometry in primary hippocampal NSCs after Samsn1 overexpression (l, m) and quantification for NSCs in the different stages of the cell cycle (n). (o, p) CCK-8 assay in primary hippocampal NSCs 0, 1, 2, 3, 4 days after plasmid transfection or virus infection. Error bars represent the . . , , . Abbreviations: Ctrl: control; Lama2-OE: Lama2-overexpression; Samsn1-OE: Samsn1-overexpression.

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