Fig 1.
Neural stem cells could proliferate and differentiate into neurons and astrocytes.
(A) Representative photomicrograph of neurospheres in culture. (B) Immunocytochemical staining of purified NSCs with Nestin. (C) Immunocytochemical staining of purified protoplasmic astrocytes with GFAP. (D) Immunocytochemical staining of purified neurons with β-tubulin-III. (E) Nucleus staining of differentiated cells from NSCs with DAPI.
Fig 2.
miR–381 promoted neural stem cells proliferation.
(A) The expression of miR–381 was measured by qRT-PCR. (B) CCK–8 was performed to detect the neural stem cells proliferation. (C) The mRNA expression of nestin was detected by qRT-PCR. (D) The protein expression of nestin was measured by Western blot. **p<0.01 and ***p<0.001.
Fig 3.
miR–381 promoted neural stem cells differentiation to neurons.
(A) Immunocytochemical staining of purified neurons with β-tubulin-III. (B) The mRNA expression of β-tubulin-III was detected by qRT-PCR. (C) The protein expression of β-tubulin-III was measured by Western blot.***p<0.001.
Fig 4.
miR–381 inhibited neural stem cells differentiation to astrocytes.
(A) Immunocytochemical staining of purified protoplasmic astrocytes with GFAP. (B) The mRNA expression of GFAP was detected by qRT-PCR. (C) The protein expression of GFAP was measured by Western blot.***p<0.001.
Fig 5.
Hes1 was the direct target of miR–381 in neural stem cells.
(A) Hes1 was predicted to be target gene ofmiR–381 by TargetScan. (B) Luciferase reporter assay was done to confirm the predictions in neural stem cells. (C) The protein expression of Hes1 was measured by Western blot in neural stem cells.***p<0.001.
Fig 6.
miR–381 promoted neural stem cells proliferation and differentiation to neurons by targeting Hes1.
(A) The protein expression of Hes1 was measured by Western blot in neural stem cells. (B) CCK–8 was performed to detect the neural stem cells proliferation. (C) The mRNA expression of Hes1 was measured by qRT-PCR in neural stem cells. (D) The protein expression of Hes1 was measured by Western blot in neural stem cells. (E) The mRNA expression of β-tubulin-III was measured by qRT-PCR in neural stem cells. (F)The protein expression of β-tubulin-III was measured by Western blot in neural stem cells. (G) The mRNA expression of GFAP was measured by qRT-PCR in neural stem cells. (F)The protein expression of GFAP was measured by Western blot in neural stem cells.**p<0.01 and ***p<0.001.