FIGURE 5:

PRMT 1 and 10 are enhanced in regenerating and resorbing flagella. (A) Immuno­blots of full-length, resorbing, and regenerating flagella using the antibodies indicated. The concentration of each sample was adjusted to 1 mg/ml, and equal volumes of sample were loaded. Tubulin stained with Ponceau S is shown as a loading control. The PRMT 1 band migrates more slowly in samples of regenerating flagella. PRMT 10 always appears as a doublet, and the upper band becomes more pronounced relative to the lower band in regenerating flagella. The amount of IFT88 is greatly increased in both resorbing and regenerating flagella, whereas the amount of KAP-GFP was only observed to increase in regenerating flagella. This is likely because the KAP-GFP lane has been underexposed so as not to overexpose the signal from IFT88. (B) Samples of full-length, resorbing, and regenerating flagella were fractionated into the freeze-thaw supernatant (FThaw), NP-40 extract (membrane fraction), and axonemes, adjusted to 1 mg/ml, separated by SDS–PAGE, and blotted, and the blot was probed with the indicated antibodies. Changes in band intensities as noted in Results are indicated with red boxes.