GSK3ß is a positive effector downstream of WNK-OSR1.

<p>(A) Gene expression determined by RT-PCR or quantitative RT-PCR analysis was examined in Neuro2A cells overexpressing GSK3ß or GSK3ß ^K85M. The value obtained from each sample was normalized to that of <i>GAPDH</i>. The value of <i>Lhx8</i> from overexpressing GSK3ß (lane 2) was set to 100. (B) Gene expression detemined by RT-PCR or quantitative RT-PCR analysis was examined in Neuro2A cells overexpressing hWNK1 under <i>GSK3ß</i> knockdown using siRNA. The value obtained from each sample was normalized to that <i>GAPDH</i>. The value of <i>Lhx8</i> from overexpressing hWNK1 (lane 2) was set to 100. (C) Gene expression determined by RT-PCR or quantitative RT-PCR analysis was examined in Neuro2A cells stimulated by retinoic acid (RA) or overexpressing GSK3ß under both <i>Wnk1</i> and <i>Wnk4</i> knockdown using siRNA. The value obtained from each sample was normalized to that of <i>GAPDH</i>. The value of <i>Lhx8</i> from overexpressing GSK3ß (lane 3) was set to 100. (D) Gene expression determined by RT-PCR or quantitative RT-PCR analysis was examined in Neuro2A cells overexpressing OSR1 or OSR1^S324D (constitutively active form of OSR1) under <i>GSK3ß</i> knockdown using siRNA. The value obtained from each sample was normalized to that of <i>GAPDH</i>. The value of <i>Lhx8</i> from overexpressing OSR1 (lane 3) was set to 100. (E) Gene expression determined by RT-PCR or quantitative RT-PCR analysis was examined in Neuro2A cells overexpressing hWNK1 or GSK3ß under <i>Osr1</i> knockdown using siRNA. The value obtained from each sample was normalized to that of <i>GAPDH</i>. The value of <i>Lhx8</i> from overexpressing hWNK1 (lane 3) was set to 100.