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. Author manuscript; available in PMC: 2014 Dec 1.
Published in final edited form as: Biochim Biophys Acta. 2013 Jul 18;1833(12):10.1016/j.bbamcr.2013.07.002. doi: 10.1016/j.bbamcr.2013.07.002

Figure 4.

Figure 4

Effect of RNA interference-mediated silencing of Sp1, Sp3, or Sp4 on the expression of the NMDA receptor subunit genes. (A) Real-time PCR revealed a down-regulation of Sp1, Sp3, and Sp4 transcripts in N2a cells transfected with Sp1, Sp3, and Sp4 shRNA, respectively. N = 6. (B) mRNA levels of Grin1, Grin2a, and Grin2b were decreased with Sp4 shRNA but not with Sp3 or Sp1 shRNA. N = 6. (C–D) Western blots revealed a down-regulation of Sp4, Sp3, and Sp1 proteins in Sp4, Sp3, and Sp1 shRNA-transfected N2a cells, respectively. β-actin served as loading control and a representative blot is shown. N = 3. (E–F) Silencing of Sp4 reduced the protein levels of GluN1, GluN2A, and GluN2B, whereas silencing of Sp1 and Sp3 did not significantly change these subunit levels. β-actin served as a loading control. N = 3. (G) Primary neurons transfected with Sp1, Sp3, or Sp4 shRNA showed decreases in Sp1, Sp3, and Sp4 transcript, respectively. N = 3. (H) mRNA levels of Grin1, Grin2a, and Grin2b were decreased with Sp4 shRNA but not with Sp3 or Sp1 shRNA in primary neurons. Grin2c levels did not decrease with Sp1, Sp3, or Sp4 shRNA in primary neurons. N = 3. *= P < 0.01, **= P < 0.01, and ***= P < 0.001.