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. 2015 Aug 4;125(9):3433–3448. doi: 10.1172/JCI75245

Figure 4. Exons 11 and 12 destabilize WNK1 in a PY motif–dependent manner.

Figure 4

(A) Schematic comparing Myc-tagged WT full-length L-WNK1, PY motif–null L-WNK1 (Y829A/Y945A), and a previously reported natural L-WNK1 splice variant lacking exons 11 and 12 (Δ11/12). (B) Steady-state abundance of full-length L-WNK1 was less than the PY-null mutant or L-WNK1 Δ11/12. n = 6; *P < 0.001 or †P < 0.01 by 1-way ANOVA, Dunnett’s post hoc comparison to full-length L-WNK1. (C) Stability of Myc-tagged L-WNK1 cDNAs in HEK-293T cells. Cells transiently expressing full-length L-WNK1, PY motif–null L-WNK1, or the Δ11/12 splice variant were treated with CHX (100 μg/ml) 24 hours after transfection and chased for 4 hours. Right panel, plot of L-WNK1 abundance during the time course, expressed as a percentage of the starting material at time zero. n = 4; *P < 0.05 by 1-way ANOVA, Dunnett’s post hoc comparison of either the PY motif–null mutant or the Δ11/12 splice variant to full-length L-WNK1 at the 4-hour time point.