Files in this Data Supplement:
Supplemental Figure S1. Northern blot of steady state testicular RNA levels from wild-type homozygote (+/+), wild-type/KO heterozygote (+/-), wild-type homozygote plus transgene (+/+R), wild-type/KO heterozygote plus transgene (+/-R), Ppp1cc -/- rescue (-/-R), and KO (-/-). The probe used is specific for Ppp1cc2 RNA. It is noteworthy that the Ppp1cc -/- rescue testis expresses about as much Ppp1cc2 RNA as does the Ppp1cc +/- testis. Note that transgenic Ppp1cc2 RNA is smaller than endogenous RNA, and truncated Ppp1cc -/- RNA is even smaller.
Supplemental Figure S2.Transmission electron micrographs of testicular rescue spermatids and sperm. A) Section of step 7 rescue spermatid showing normal development of acrosomal vesicle (arrow). B) Transverse section showing normally developed portion of midpiece of testicular rescue sperm. C) Transverse (upper arrow) and sagittal (lower arrows) sections through midpieces of testicular rescue sperm. Note the eccentric position of the outer dense fibers and axoneme within the encircling mitochondria in the transverse section, and the nonuniformity of mitochondrial gyre morphology and spacing as indicated by the arrows in the sagittal section. D) Another example of nonuniform mitochondrial gyre morphology and spacing, creating potential weak spots (arrows) in the testicular rescue sperm midpiece. E, F) Sagittal sections through the midpieces of testicular wild-type sperm showing uniform mitochondrial gyre morphology and spacing. G, H) Condensing chromatin within normally developing sperm heads of late elongating spermatids from rescue mice. I) Transverse section through principal piece of testicular rescue sperm showing malformed (gapped) fibrous sheath rib (arrows). Note that ODFs 3 and 8 have not yet completely disappeared indicating that this section of the principal piece is near or at the midpiece-principal piece junction. J) Sagittal section through principal piece of testicular rescue sperm near the midpiece-principal piece junction. Arrow points to unusually large gap between fibrous sheath ribs, creating a potential weakspot through which ODFs and attached axonemal doublets can extrude. Note that in this case, the distal mitochondrial sheath abuts the annulus (arrow heads) and the principal piece, as in wild-type sperm. Original magnification A ×35,000; B ×60,000; C, D, and E ×20,000; F ×28,000; G ×15,000; H ×20,000; I ×70,000; and J ×40,000."