Figure 8.
Hemicentin is not required for later maintenance of the utse–seam B-LINK. (A) A diagram detailing timing of RNAi and utse–seam connection analysis in B–D. (B) The percent of animals ruptured after 120 h on control (T444T empty vector) or hemicentin RNAi that was started at either the early-L4 (control n = 33, hemicentin n = 26), the mid-L4 (control n = 25, hemicentin n = 40), the late-L4 (control n = 26, hemicentin n = 48), or the young adult stage (control n = 243, hemicentin n = 127). ***P < 0.001, **P < 0.01, *P < 0.05, Fisher’s Exact Tests. (C) Ventral fluorescence images of the utse (magenta) and seam (green) on hemicentin RNAi started at either the L1 stage or at the young adult (YA) stage (L1 data shown previously in Fig. 7, B and C). Regions where the utse has pulled away from the seam (utse–seam gaps) indicating defects in the B-LINK are highlighted with white dotted lines and quantified on the right (n = 10 each). ***P < 0.001, unpaired two-tailed Student’s t tests. (D) Ventral fluorescence images of the utse and seam on fibulin-1 RNAi started at the L1 stage or at the mid-to-late-L4 stage (L1 data shown previously in Fig. 7, B and C). Utse–seam gaps (dotted lines) indicating defects in the B-LINK are quantified on the right (n = 10 each). *P < 0.05, unpaired two-tailed Student’s t tests. (E) A schematic detailing timing of α1-type IV collagen RNAi in F. (F) Quantification of α1-type IV collagen::mRuby2 levels at the B-LINK when Rup phenotypes began after L1-initiated α1-type IV collagen RNAi compared to a1-type IV collagen::mRuby2 levels at the B-LINK after 120 h on α1-type IV collagen RNAi started at the early-L4 stage. Error bars represent SEM. ****P < 0.0001, unpaired two-tailed Student’s t test. Box edges in boxplots depict the 25th and 75th percentiles, the line in the box indicates the median value, and whiskers mark the minimum and maximum values. Scale bars, 10 µm.