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Fig. 5 | Molecular Medicine

Fig. 5

From: Normal Human Epidermal Keratinocytes Express In Vitro Specific Molecular Forms of (Pro)Filaggrin Recognized by Rheumatoid Arthritis-Associated Antifilaggrin Autoantibodies

Fig. 5

Immunoblotting reactivity of RA serum and MAbs AHF1 and AHF7 on urea extracts of cultured keratinocytes and of normal human epidermis separated by two-dimensional nonequilibrium pH gel electrophoresis/SDS-PAGE

In the extracts from cultured keratinocytes (upper half), the RA serum and the AHF MAbs detect a diffuse protein spot (migration-like pI markers ranging from 5.4 to 5.9 and molecular weight above 100 kDa). AHF1 detects an additional group of molecules with lower molecular weight and more basic pI (ranging from 70 to 100 kDa and from pI 6 to 7). In the extracts from normal epidermis (lower half), AHF1 detects profilaggrin (molecular weight above 200 kDa and pI from 5.4 to 5.9), comma-shaped filaggrin (molecular weight of 37–40 kDa and pI from 6 to 9) and a series of molecules of intermediate size and charge. AHF7 detects a protein doublet corresponding to the most acidic part of the urea-soluble filaggrin. Like AHF7, the RA serum weakly detects the most acidic part of the urea-soluble filaggrin (arrowhead) and recognizes the same intermediate molecules between filaggrin and profilaggrin like AHF1. Molecular weight markers are on the left and migration of pI markers are on the top of each membrane. NEpHGE, nonequilibrium pH gel electrophoresis.

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