Journal: bioRxiv

Article Title: A spinal synergy of excitatory and inhibitory neurons coordinates ipsilateral body movements

doi: 10.1101/2023.03.21.533603

Figure Lengend Snippet: (A) Whole-mount β-galactosidase staining on E10.5 and E11.5 Hes2 iCre ;R26 LSL-LacZ embryos to characterize the expression of Hes2 iCre . (B) Cross-sections of E12.5 lumbar spinal cord from Hes2 iCre ;Gata3 nlsLacZ ;R26 LSL-Sun1-GFP embryos stained with antibodies for GFP to label Hes2 neurons (green), β-gal ( Gata3 nlsLacZ ) to label V2b neurons (red), and Chx10 to label V2a neurons (blue). Scale bar: 100 μm. (C) Bar graph showing the percentages of V2a (Chx10+) and V2b ( Gata3 nlsLacZ +) neurons within the Hes2 lineage (Hes2+). (D,E) Bar graphs showing the percentage of V2a (Chx10+) (D) and V2b ( Gata3 nlsLacZ +) (E) neurons captured by the Hes2 iCre . (F) Cross-sections of E12.5 lumbar spinal cord from Hes2 iCre ;Gata3 nlsLacZ ;R26 LSL-Sun1-GFP embryos either heterozygous (left) or homozygous (right) for the iCre allele. The sections were stained with antibodies for GFP (green), β-gal ( Gata3 nlsLacZ ) (red) and Chx10 (blue). Scale bars: 100 μm. (G) Bar graph showing the absence of changes in the number of Hes2 neurons in Hes2 knockout embryos compared to heterozygous littermates, assessed by two-tailed Student’s t-test.

Article Snippet: The following primary antibodies were used: guinea pig anti-Chx10 (1:2000) , rabbit anti-βgal (1: 5000; Cappel), rabbit anti-HB9 (1:6000) , goat anti-GFP (1:1000; Rockland).

Techniques: Staining, Expressing, Knock-Out, Two Tailed Test

Journal: BMC Developmental Biology

Article Title: Analysis of Thisbe and Pyramus functional domains reveals evidence for cleavage of Drosophila FGFs

doi: 10.1186/1471-213X-10-83

Figure Lengend Snippet: Restricting the source of full-length and truncated Ths and Pyr constructs reveals a functional difference . (A-F) Immunohistochemistry on stage 11 embryos, lateral view, all constructs driven with ZenKr-GAL4; embryos were stained using an anti-Eve antibody. (A) White numbers indicate position of numbered Eve-positive clusters; ZenKr-GAL4 supports expression in clusters 4-7. (A-F) Eve staining reveals additional Eve-positive cells outside the ZenKr domain for (A) HA-Ths-Myc (B) HA-Ths 1-158 (C) HA-Ths 1-403 (D) HA-Pyr-Myc (E) HA-Pyr 1-348 (F) HA-Pyr 1-466 . (G,H,I) Eve-positive cells per cluster were counted in each hemisegment for 25 embryos per construct tested and averaged. Error bars indicate standard error. (G) The hatched line at "3" represents the wild-type level of Eve-positive cells. The gray box represents the source of expression supported by ZenKr-GAL4. Plot of Eve-positive cells generated by ZenKr-GAL4 → pUASt-HA-Ths-Myc as compared to ZenKr-GAL4 → pUASt-HA-Pyr-Myc shows that Pyr has greater functional activity than Ths. Ths and Pyr both give a graded output of Eve-positive cells with the most cells in the source domain. (G') ZenKr-GAL4 driving UAS-lacZ and stained with anti-βgal shows the domain of the driver in the posterior dorsal ectoderm of the embryo. (H) ZenKr-GAL4 → pUASt-HA-Ths 1-158 does not have the same Eve-positive profile, instead it results in more Eve-positive cells in clusters 8-11. ZenKr-GAL4 → pUASt-HA-Ths 1-403 has increased activity locally but similar levels of function to HA-Ths-Myc at long-range (I) ZenKr-GAL4 → pUASt-HA-Pyr 1-348 and ZenKr-GAL4 → pUASt-HA-Pyr 1-466 both retain a graded profile of Eve-positive cells, although HA-Pyr 1-348 supports more Eve-positive cells in distant clusters 8-11 as compared to HA.-Pyr 1-466 .

Article Snippet: Primary antibodies used were: anti-Even skipped rabbit (1:1000, M. Frasch) and anti-βgal rabbit (1:250, Molecular Probes).

Techniques: Construct, Functional Assay, Immunohistochemistry, Staining, Expressing, Generated, Activity Assay