Hematopoietic Protein-1 Regulates the Actin Membrane Skeleton and Membrane Stability in Murine Erythrocytes
Figure 3
Expression of WAVE complex mRNA and protein in wildtype (WT) and Hem1−/− mouse erythrocytes.
(A) (Left) Real-time PCR of FAC-sorted erythroblasts from WT mice shows relative expression of Hem1 and WAVE complex (Abi1, Abi2, Sra1, and Wave2) mRNA normalized to β-actin. Expression of Hem1 mRNA is shown relative to kidney mRNA. Expression of WAVE component mRNAs are shown relative to a CT value of 38, which represents an arbitrary low expressing tissue. (*), p<0.0007; (**), p<0.0004; (***), p<0.0001. n = 3 mice per genotype. (Right) Expression of WAVE complex mRNA are shown in Hem1 erythroblasts versus WT erythroblasts. Abi2 p<0.0004 (B) Immunoblots of purified erythrocyte ghosts from WT and Hem1−/− mice, loaded according to equivalent levels of β-actin. Each lane is representative of at least 3 individuals per genotype. Hem-1 protein (upper band, 110 kDa) is not detectable in erythrocyte ghosts from Hem1−/− mice. Although Abi2 mRNA levels are reduced in Hem1−/− erythrocytes, Abi2 protein levels are relatively normal, likely due to post-transcriptional and/or post-translational regulation. Numbers below each scan represent protein expression levels in Hem1−/− relative to WT samples. Samples were loaded based on equivalent total protein.