Larger Image FIGURE 2. Rab7 knockdown resulted in loss LRO sensor tissue. (A) Experimental setup for the shown experiments: Injections at four-cell stage (left) targeted the dorsal/dorsolateral mesoderm to analyze superficial mesoderm induction (foxj1; middle) or somitic mesoderm specification (myf5; middle) at gastrula stages, following analyses of general LRO formation (tekt2; right) or sLRO specification (dand5/nodal1; right) during neurulation. (B) Wild type crescentic foxj1 transcripts were (C) unaltered or (D) mildly affected by low dose rab7 TBMO injections (black outlined arrowheads). (E) Quantification of foxj1 expression. (F) Expression domain of tekt2 in the LRO of st. 17 embryos, (G) depicted slightly stronger posterior and reduced anterior signals upon low dose rab7 TBMO treatment. (H) Wild type dorsolateral myf5 expression (I) was significantly reduced in rab7-morphant embryos (black outlined arrowheads). (J) Quantification of myf5 expression. (K,O) Expression of dand5 and nodal1 in the somitic domains of the LRO of control and (L,P) coMO injected specimen, respectively. (M) Transcripts of dand5 or (Q) nodal1 were lost (black arrowheads) or severely reduced (black outlined arrowheads) in rab7-deficient embryos (low dose TBMO). (N,R) Quantification of dand5 and nodal1 expression, respectively. (S) Left-sided unilateral knockdown (marked by asterisk) of low dose rab7 TBMO in myod1 expressing LRO, (S′) section revealed lost superficial expression of myod1 at injection side (black arrowhead). cLRO, central LRO; co, control; coMO, control Morpholino oligomere; d, dorsal; l, left; LRO, left-right organizer; N, number of experiments; n, number of evaluated embryos; n.s., not significant; r, right; sLRO, somitic LRO; st., stage; TBMO, translation blocking Morpholino oligomere; v, ventral; wt, wild type. Image published in: Kreis J et al. (2022) Copyright © 2022. Creative Commons Attribution license Gene Synonyms Species Stage(s) Tissue foxj1.L foxj1.1, foxj1-a, foxj1-b, hfh-4, hfh4, xfoxj1 X. laevis Throughout NF stage 10 mesoderm dorsal marginal zone myf5.L myf-5, Xmyf-5, Xmyf5 X. laevis Throughout NF stage 11 mesoderm dorso-lateral marginal zone tekt2.L tekt2-t, Tektin-t X. laevis Throughout NF stage 17 left-right organizer gastrocoel roof plate dand5.L 51-B6, cer2, cerl2, charon, cktsf1b3, coco, crl2, dante, grem3, S10-51-B6, xCoco X. laevis Throughout NF stage 17 left-right organizer gastrocoel roof plate myod1.L mf25, MyoD, myod1-a, myod1-b, myogenic factor 3, XMyoD, XtmyoD X. laevis Throughout NF stage 17 presomitic mesoderm nodal1.L nodal, nodal-related 1, nr1, nr1-a, spaw, Xnr-1, Xnr1 X. laevis Throughout NF stage 17 left-right organizer gastrocoel roof plate Image source: Published Experiment + Assay Source Phenotypes and Disease Xla Wt + rab7a MO + NF10-11 (in situ hybridization) Fig. 2 B-G Expression Phenotype decreased amount foxj1.L expression in dorsal marginal zone mislocalised tekt2.L expression in left-right organizer Xla Wt + rab7a MO + NF10-11 (in situ hybridization) Fig 2 H-J Expression Phenotype decreased amount myf5.L expression in dorso-lateral marginal zone decreased amount myf5.L expression in presomitic mesoderm Xla Wt + rab7a MO + NF17 (in situ hybridization) Fig. 2 K-R Expression Phenotype decreased amount dand5.L expression in left-right organizer absent dand5.L expression in left-right organizer absent nodal1.L expression in left-right organizer decreased amount nodal1.L expression in left-right organizer Xla Wt + rab7a MO + NF17 (in situ hybridization) FIG 2 S' Expression Phenotype decreased amount myod1.L expression in epithelial cell Permanent Image Page Printer Friendly View XB-IMG-195318