Stage vulval cells. (A92L9) Corresponding GFP fluorescence photomicrographs. (A2D, A9-D9) egl-17::gfp (ayIs4); (E2F, E92F9) zmp-1::gfp (syIs49); (G, H, G9, H’) ceh-2::gfp (syIs54); (I, J, I9, J9) daf-6::yfp (bhEx53) and (K2L, K92L9) cdh-3::gfp (syIs50). The expression patterns of all markers are impacted in hda-1 animals. Arrows mark the center of vulval invagination. B1, B2, C, D, E, and F refer towards the presumptive vulval cell fates vulB1, vulB2, vulC, vulD, vulE, and vulF, respectively. Scale bar is 10 mm.undergoing morphogenetic modifications (Gupta et al. 2003). The fos-1 locus encodes three transcripts which have some functional variations. fos-1a (syIs123 fos-1a::yfp) is virtually exclusively present within the AC and is definitely the target of hda-1 throughout AC invasion (Matus et al. 2010). fos-1b(syIs137 fos-1b::cfp) is observed at a low level in many uterine cells, like the AC (Sherwood et al. 2005), and itdoes not seem to play a role in AC invasion. Yet another fos-1 transcript, fos-1c, is expressed in uterine p lineage cells and involved in utse formation (Oommen and Newman 2007). We examined syIs123 and syIs137 transgenic animals and discovered that while fos-1a::yfp was undetectable in vulval cells for the duration of the L3 and L4 stages (data not shown), fos-1b::cfp was expressed inside a subset of vulval progeny.n Table two Vulval cell fate specification defects in hda-1 RNAi animals Cell Fate Marker zmp-1::gfp ceh-2::gfp egl-17::gfp cdh-3::gfp daf-6::yfp RNAi A L4440 hda-1 L4440 hda-1 L4440 hda-1 L4440 hda-1 L4440 hda-1 one hundred ND one hundred 81 one hundred 60 one hundred 66.six 100 60 100 100 B1/2 Vulval Cell Form C one hundred 83.3 D one hundred 100 E one hundred 62.five F n 50 24 50 27 50 30 50 15 50100 66.six 100 76.6100 40 100 83.Digitoxigenin MedChemExpress 3Percentage of vulval cells obtaining GFP fluorescence are shown.Apiin NO Synthase A, C, D, E, and F refer towards the presumptive vulval cell fates vulA, vulC, vulD, vulE, and vulF, respectively. vulB1, and vulB2 are listed collectively as B1/2. L4440 refers to control RNAi animals. RNAi, RNA interference; n, number of animals examined; ND, not accomplished.Volume three August 2013 |Role of hda-1 in Caenorhabditis elegans |Figure 4 hda-1 expression within the vulva and AC. (A2E) sEx13706 and (F) bhEx68. (A, B) Pn.px cells. (C, D) Pn.pxx cells. (E, F) Pn.pxxx cells. Triangles mark the center of vulval invagination. The presumptive vulval cell types A (vulA), B (vulB1 or vulB2), and D (vulD) are shown. The AC is shown with arrows. In (B), P5.p is in the procedure of dividing and has lowered degree of GFP fluorescence. Scale bar is ten mm. Figure 3 lin-11 and fos-1 expression is altered in hda-1 mutants. DIC and corresponding fluorescent images of animals expressing a translational fos-1::cfp reporter. (A and B) Control L4440 RNAi-treated midL4 animal showing fos-1 expression in presumptive vulD cells.PMID:23310954 (C2H) mid/late-L4 stage animals showing fos-1 expression in presumptive vulD, vulE and vulF cells. (I, J) hda-1 knockdown causes reduction in fos-1::cfp expression. Diffuse CFP fluorescence is observed within the area overlapping with presumptive vulD cells. lin-11 expression is detected in vulval cells in handle RNAi-treated animals (K, L) but is absent in hda-1-RNAi treated animals (M, N). A few of the GFP fluorescing cells are marked by arrowheads and arrows (D, E and F refer to vulD, vulE and vulF, respectively). mL4: mid-L4, lL4: late-L4. Asterisk in panel N points to VC neuronal cells. Scale bar is 10 mm.handle, n = 25) (Figure 3, I and J). The pattern was comparable in late-L4 animals (information not shown). These resul.
