Fig. 5

Depletion of Yy1 in primary cortical neurons reduces GluR1 protein levels. (a) Western blot from primary neurons after knockdown of Yy1 with siRNA (siYy1). Primary cortical neurons were transfected with scramble (Scr) or siYy1. Whole cell extracts were prepared and subjected to immunoblot with anti-GluR1, anti-Yy1, and anti-Actin antibodies. (b) qRT-PCR shows that Yy1 and Senp1 mRNA levels are statistically decreased following siYy1, while GluR1 mRNA levels are not statistically decreased. Graphs indicate three independent biological replicates. Error bars represent one standard deviation from the mean. **(p < 0.01); *(p < 0.05); ns. not significant (p > 0.05). p value was determined using two-tailed unpaired t test. (c) Anti-GluR1 immunostaining of the shRNA transfected cells. Primary cortical neurons were transfected with shRNA Control (shCtrl), shYy1–2, or shYy1–3. GFP encoded in the shRNA vector tracks the transfected cells. Scale bar: 10 μM. (d) Quantification of GluR1 level in control and shYy1 transfected neurons. The mean intensity of GluR1 signals was determined using Image J software. *** (p < 0.0001). p value was determined using two-tailed unpaired t test