FIG. 7.

Activation of crystallin expression by MafA requires the integrity of serines 14 and 65. Chicken embryo NR cells were transfected with empty RCAS (RCAS), RCAS-MafA (WT), or RCAS-S14A/S65A (S14A/S65A) plasmids. (A) Transcription of αA-, βB1-, and δ1-crystallin genes was analyzed 6 days after transfection by RT-PCR. MafA- and ribosomal s17-specific PCR were performed as controls. (B) Cellular lysates prepared 6 days after transfection were analyzed by Western blotting with α-crystallin-, β-crystallin-, δ-crystallin-, or MafA-directed sera. Relative amounts of proteins expressed in the WT versus S14A/S65A MafA-containing cultures were determined according to densitometry of autoradiograms and are indicated on the right. A β-actin Western blot was performed as a loading control.