Fig. 5.

CDK-11.1 is required for full activation of the RAS-ERK pathway. (A) Dissected germ lines stained for DNA (blue), activated ERK (dpMPK-1, green), and CDK-11.2 (red). In wild-type germ lines, dpMPK-1 levels peak in late pachytene (green dashed line) and again in diakinesis (red dashed line), The fog-2(q71) germ lines serve as a negative control where very little or no dpMPK-1 staining is observed. cdk-11.1(tm5495) mutant hermaphrodites display decreased levels of dpMPK-1 in both late pachytene and diakinesis, while cdk-11.2(bs101) animals exhibit a similar reduction in the level of dpMPK-1 in late pachytene but a milder reduction during diakinesis. Note that cdk-11.1(tm5495) mutants show an increase in the level of CDK-11.2 throughout the entire germ line when compared to wild-type germ lines. Scale bars are 25 μm in all images. B and C) Quantitation of dpMPK-1 levels (plotted as arbitrary units, A.U.) during late pachytene and diakinesis in wild-type, cdk-11.1(tm5495), cdk-11.2(bs101) and fog-2(q71) germ lines. Black bars represent the mean. A one-tailed t-test was used to determine if mutants exhibit a significantly different dpMPK-1 level relative to the wild type (*p < 0.05,**p < 0.0001, ***p < 0.00001). The number of gonads scored (late pachytene/diakinesis) = 29/31 for wild type, 22/23 for cdk-11.1(tm5495), 14/13 for cdk-11.2(bs101), and 8/10 for fog-2(q71). (D) Quantitative immunoblot demonstrating that loss of CDK-11.1 leads to an increase in expression of CDK-11.2. Whole worm lysates were prepared and the equivalent of thirty adult animals were loaded in each lane.