Table 3.

Western blot of full-length MLP-1 + mCherry (lane F), NH₂-terminal 112-amino-acid fragment + mCherry (lane N), and COOH-terminal 118-amino-acid fragment + mCherry (lane C)

mCherry Ab
Lane	Mol Mass, kDa	Comparison Between Measured and Calculated Molecular Masses
F	75.5	Lane F, band 1 (top): MLP-1 weighs 20.17 kDa; runs at 47.8 kDa + mCherry 76.6 kDa. Compare with measured 75.5 kDa. Conclusion: full length + mCherry. Detected by mCherry Ab, NH2-terminal MLP-1 Ab, and COOH-terminal MLP-1 Ab.
N	50.5	Lane N, band 1: NH2-terminal fragment is 112 residues. Mol mass 11.5 kDa corrected for effector domain (time 2.37) = 27.3 kDa + mCherry = 56.1 kDa. Compare with measured 50.5 kDa. Conclusion: full-length NH2-terminal fragment + mCherry. Detected by mCherry Ab and NH2-terminal MLP-1 Ab but not COOH-terminal MLP-1 Ab.
C	59.0	Lane C, band 1: COOH-terminal fragment is 118 amino acids. Mol mass 12.2 kDa; corrected for effector domain should run at 28.9 kDa + mCherry = 57.7 kDa. Compare with measured 59.0 kDa. Conclusion: full-length COOH-terminal fragment with mCherry. Detected by COOH-terminal MLP-1 Ab and mCherry Ab.
	43.0	Lane C, band 2: alternative start site protein might run at 6.8 kDa; corrected to 16.1 kDa + mCherry 44.9 kDa. Compare with measured 43.0 kDa. Detected by COOH-terminal MLP-1 Ab and mCherry Ab.

Blotted with anti-mCherry (as in Fig. 7). Ab, antibody; MLP-1, myristoylated alanine-rich protein kinase C substrate-like protein-1.