Figure 1. Characterization of N- and C-terminal endogenously tagged TDPT-1 strains:(A-B) Live confocal imaging of the wrmScarlet::
tdpt-1 strain. Top panels show wrmScarlet::TDPT-1 alone, the middle panels show the DIC image alone, and the lower panels show the merged images captured at 40X. Scale bars represent 25 μm. (A) Localization of wrmScarlet::TDPT-1 in the germ line of the hermaphrodite gonad. The white dashed line indicates a break/boundary of two images from the same worm that do not align perfectly. The spermatheca is indicated by white arrows. (B) Localization of wrmScarlet::TDPT-1 in somatic cells of the worm head. (C) Localization of wrmScarlet::TDPT-1 in embryos. (D) Percent viable progeny of tagged TDPT-1 strains compared to wild type (N2). The progeny of at least 30 hermaphrodites were scored for each genotype. The total number of progeny scored for each genotype was as follows: N2= 6,779; wrmScarlet::
tdpt-1= 8,204;
tdpt-1::3xflag= 7,149. Statistical analysis was performed using Student's t-test, n.s. indicates not significant. (E) Western blot of TDPT-1 protein levels in wild type (N2) versus TDPT-1::3xFLAG. The TDPT-1::3XFLAG band (~44 kDa) is observed in lysates from the
tdpt-1::3xflag strain and absent in wild type (N2) lysates. α Tubulin (50 kDa) control bands are present in both strains.