Figure 6.

SKP1 increases FBXO45 binding to NMNAT2. A, coIP from 293 cells shows GFP–FBXO45 binds NMNAT2–MYC, and GFP–SKP1 fails to bind NMNAT2–MYC. B, coIP showing HA–SKP1 will bind NMNAT2–MYC in the presence of FLAG–FBXO45. C, coexpression of HA–SKP1 increases binding of FLAG–FBXO45 to NMNAT2–MYC. D, quantitation, in arbitrary units (A.U.), of coIP between FLAG–FBXO45 and NMNAT2–MYC in the presence or absence of HA–SKP1. Significant increase in FLAG–FBXO45 coIP with NMNAT2–MYC occurs when HA–SKP1 is present. E, summary showing SKP1 increases NMNAT2 binding to FBXO45. B and C, shown are representatives of at least three independent experiments. D, quantitation was done with a minimum of three replicates. Significance was determined using Student's t test, and error bars represent standard error of mean. ***, p < 0.001; IP, immunoprecipitation.