Xenbase Image ID: 139010

FIGURE 3. Knockdown of Hspa5 inhibits pronephros formation. A–C, phenotype of Xenopus tadpoles injected with Hspa5MO2 into one ventral-vegetal blastomere at the eight-cell stage. Either 15 or 30 ng of Hspa5MO2 was co-injected with LacZ mRNA into one of the ventral-vegetal blastomeres of eight-cell stage embryos. A, anterior-dorsal view of mildly affected embryos at stage 16. B, lateral view of mildly affected embryos at stage 32. LacZ staining indicates the injected side; the uninjected side was used as a negative control. C, phenotype categorization of embryos injected with 15 or 30 ng Hspa5MO2. Over 90% of the embryos showed a normal or mild repression of anterior axis development after the injection. The severe phenotype refers to failure of gastrulation. D–F , depletion of Hspa5 inhibits expression of pan-pronephros marker genes such as pax2, lhx1, and atp1b1. Eight-cell stage embryos were injected with LacZmRNA (100 pg) into one of the vegetal-ventral blastomeres as a lineage tracer along with Hspa5MO2 (30 ng/embryo). The injected embryos were collected at stage 32, and the expression of pronephric marker genes as indicated was examined by whole mount in situ hybridization. D, E, and F, the uninjected side. D , E , and F , expression of pax2, lhx1, and atp1b1 on the MO-injected side of the same embryos, respectively. D , E , and F , transverse sections of embryos in D, E, and F accordingly. G–H , expression of smp30 (tubule) and nephrin (glomus) at the Hspa5MO2-injected side was inhibited. G and H, the uninjected side. G and H , Hspa5MO2-injected side. G and H , transverse sections of embryos shown in G and H, respectively. I–L , expression of pax2, lhx1, smp30, and nephrin in the tadpoles injected with 30 ng of standard control MO/embryo with 100 pg of LacZ mRNA into one vegetal-ventral blastomere at the eight-cell stage. I–L, uninjected side. I -–L , standard control MO-injected side (CMO-lacZ). Image published in: Shi W et al. (2015) Copyright © 2015. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Gene Synonyms Species Stage(s) Tissue pax2.L LOC108697493, pax-2, pax2-a, pax2-b, XPax-2, XPax2 X. laevis Throughout NF stage 32 pronephric tubule pronephric nephrostome lhx1.L lim-1, Lim1, lim1, Xlim-1, Xlim1 X. laevis Throughout NF stage 32 pronephric mesenchyme atp1b1.S atp1b1a, Beta1-Na/K-ATPase, Na+-K+-ATPase, β1-NaK-ATPase X. laevis Throughout NF stage 32 pronephric kidney pronephric nephron pronephric tubule visceral pouch nphs1.S nephrin X. laevis Throughout NF stage 32 pronephric kidney pronephric mesenchyme glomus rgn.L regucalcin, smp30, xsmp-30, xsmp30 X. laevis Throughout NF stage 32 pronephric kidney pronephric mesenchyme Image source: Published Larger Image Printer Friendly View

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