Microscopically normal training condition observed NSCs clouds， free， loose cells grow bigger， quantity is less， the ball more， and hypoxia condition， the quantity is less， the cells NSCs loose， small Immediately after inoculation difference in the microscope， a large， round cell borders， refractive After a 24h cell formation， irregular cell suspension mass， some cells， appear more cellular debris， with the passage of time， most of the cells suspended unceasingly split growth， by a number of cells with small group of cells， gradually grew by dozens to 100 cells sizes， BrdU cells suspended NSCs showed marked immunofluorescence green Determine the MTT method for detecting oxygen， and testing time 8h NSCs conditions of U0126 join 6h cell activity， proliferation and maintain peak at this level， after joining and folic acid can promote the proliferation of inhibiting proliferation， folic Fluorescence quantitative polymerase chain reaction (PCR) ERK1/2 expression level detection and oxygen to peak， after 6h expressed in this level， maintain the untreated group of folic acid， Join U0126 6h conditions， to express， also maintain at this level， U0126 + folic acid U0126 below lack Western blot ERK1 1/2 phosphorylation method to detect the expression level， express to peak oxygen 6h， maintain at this level of untreated group， folic acid， Join U0126 6h conditions， to express， also maintain at this level， U0126 + folic acid U0126 below lack
Microscope， observed under normal culture growth of NSCs into the group， free， loose cell less， larger sphere in larger quantities， but under hypoxic conditions NSCs， the number of small， cell loose ball Immediately after inoculation， observed under phase contrast microscope， cells were large round bright， clear boundary， refraction and 24h after the formation of a large number of suspended cells， irregular cell clumps， some cells attached to the wall， there are more cell debris， over time， most of the growth of suspension cells to keep dividing by the number of cells with small cell groups， gradually grow into the tens to hundreds of cells composed of cells of different sizes suspended ballBrdU labeled fluorescence signal of NSCs into the green MTT assay to determine ischemic time of 8h， and detection by adding U0126 cell activity under the conditions of NSCs， 6h proliferation reached a peak， then maintained at this level， and the addition of folic acid can promote the proliferation and inhibition of proliferation of folate Quantitative PCR detection of ERK1 / 2 expression， the expression of reoxygenation 6h reached a peak， remaining at this level， folic acid deficiency group than in the control group; adding U0126 conditions， 6h expression reached its peak， after which remained at this level， U0126 + folic acid deficient group than U0126 Western blot， phosphorylation of ERK1 / 2 expression， The peak expression of reoxygenation 6h， and then maintained at this level， folic acid deficiency group than in the control group; adding U0126 conditions， 6h expression reached its peak， after which remained at this level， U0126 + U0126 group was lower than folate
