Figure 1.—

Wild-type and mutant protein sequences of WISP. (A) Amino acid sequence of the predicted PAP domain of WISP was aligned against those of GLD-2 proteins of C.elegans, Xenopus, humans, and mice. Identical or similar residues between the sequences are indicated with an asterisk or dot(s), respectively. (B) Schematic of wisp alleles. wisp⁴¹ and wisp²⁴⁹ have nonsense mutations in the coding region that result in truncated proteins of 323 aa and 570 aa, respectively. wisp²⁴⁸ has a point mutation at position 1152, resulting in a Ile-to-Asp change. wisp^12-3147 has point mutations at positions 577 and 1151, which result in Met-to-Val and Thr-to-Ile substitutions, respectively. (C) Western blot analysis. Ovaries were dissected from wild-type, wisp⁴¹/wisp⁴¹, wisp²⁴⁸/wisp²⁴⁸, and wisp²⁴⁹/wisp²⁴⁹ females. Ovarian protein extracts were separated by SDS–PAGE, blotted, and probed with affinity-purified polyclonal anti-WISP directed against the 411 aa C-term part of the protein or with anti-α-tubulin (loading control; TUB). A protein of ∼140 kDa is detected in ovarian extracts of wild-type and wisp²⁴⁸/wisp²⁴⁸ females. This band is absent in ovarian extracts of wisp⁴¹/wisp⁴¹ and wisp²⁴⁹/wisp²⁴⁹ females.