Validating groupbox

Posted by / 10-Jan-2018 11:49

To test the functional significance of Hmgb1 release, ischemic or normal retinas were treated with the neutralizing anti-Hmgb1 antibody or recombinant Hmgb1 protein respectively.To elucidate in which cell type Hmgb1 exerts its effect, primary retinal ganglion cell (RGC) cultures and glia RGC cocultures were treated with Hmgb1.RGCs were grown in serum-free basal media (Neurobasal/B27 media; Invitrogen, Carlsbad, CA).Mixed glial cells were prepared from the brains of neonatal (postnatal day 2) mice as previously described.) and glucose- and sodium pyruvate free basal media (Neurobasal media; Invitrogen) for 4 hours at 37°C.

Recombinant Hmgb1 (total 1 μg in 2 μL of PBS, catalog number: 1690HM; R&D Systems, Inc., Minneapolis, MN) was administered by intravitreal injection as described in Yoneda et al.Autophagy plays many physiological and pathophysiological roles.However, the roles and the regulatory mechanisms of autophagy in response to viral infections are poorly defined in teleost fishes, such as grass carp ( kidney (CIK) cells, and thereby stimulate autophagy.PVDF membrane was incubated 5 minutes in a working solution.Quantification of the protein bands was performed using image acquisition and analysis software (Quantity One; Bio-Rad Laboratories, Hercules, CA).

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