rabbit anti β h spectrin  (Developmental Studies Hybridoma Bank)

Journal: G3: Genes|Genomes|Genetics

Article Title: Transcriptomic analysis of feminizing somatic stem cells in the Drosophila testis reveals putative downstream effectors of the transcription factor Chinmo

doi: 10.1093/g3journal/jkab067

Figure Lengend Snippet: Genotype ontology and gene expression in chinmo -depleted somatic cells. (A) Scatter (Volcano) plot for genes in chinmo -depleted somatic cells ( tj>GFP+chinmo-RNAi ) compared with controls ( tj>GFP ). The x-axis is the log 2 of the fold change and the y-axis is the negative log 10 of the adjusted P -value (P adj ). Gray line indicates P adj = 0.05. Gray circles indicate genes with log 2 (FC) between −1 and 1 (corresponding to FC between 0.5 and 2). Blue circles indicate genes with log 2 (FC) greater than 1 (corresponding to FC >2). The larger black circles indicate the differentially upregulated genes β H -spectrin , pyd , scrib , and DE-cad. ( B) Gene Ontology. DAVID Functional Annotation Clustering analysis depicting enriched biological processes for the 100 upregulated genes with putative Chinmo binding sites. The enrichment score plotted on the y-axis is based on the Fisher’s exact P -values of each gene in the group. The higher the score, the more enriched is the group. Genotype: (A) w/Y; tj-Gal4, UAS-GFP nls /+; UAS-chinmo-RNAi/UAS-Dcr-2 .

Article Snippet: The following primary antibodies were used: goat anti-Vasa (1:50, dC-13, Santa Cruz), rabbit anti-Vasa (1:3000; gift of Ruth Lehmann, Whitehead Institute, MA, USA), guinea pig anti-Tj (1:5000; gift of Dorothea Godt, University of Toronto, ON, Canada), mouse anti-Fasciclin (Fas) 3 (1:50; Developmental Studies Hybridoma Bank (DSHB), rat anti-DE-cad (1:25, DSHB), mouse anti-Pyd (clone Pyd2) (1:10) (DSHB); rabbit anti-β H -spectrin (1:250) (gift of Claire Thomas, Penn State University, PA, USA); rabbit anti-GFP (1:250) (Invitrogen), mouse anti-β-gal (1:50) (DSHB).

Techniques: Gene Expression, Functional Assay, Binding Assay

Journal: G3: Genes|Genomes|Genetics

Article Title: Transcriptomic analysis of feminizing somatic stem cells in the Drosophila testis reveals putative downstream effectors of the transcription factor Chinmo

doi: 10.1093/g3journal/jkab067

Figure Lengend Snippet: chinmo -deficient somatic cells upregulate β H -Spectrin. (A–A”) xy-section of a WT ovary shows that β H -spectrin (white) is present in somatic cells. Bracket in A” indicates the niche cells. Arrows in A” indicate apical enrichment of β H -spectrin in follicle cells. (B–B”) In a WT testis, β H -spectrin (white) is expressed strongly in niche cells (outlined by a dashed line in B”) and at very low levels in other somatic cells. (C–D”) In a chinmo ST mutant testis (C–C”) or a testis somatically depleted for chinmo (D–D”), β H -spectrin (white) is ectopically expressed in feminized somatic cells. Arrows in C” and D” indicate apical enrichment of β H -spectrin in chinmo -depleted somatic cells. In (A–D), Vasa (green) marks the germline and Tj (magenta) marks cyst cells. Scale bar = 10 μm. Time point in (A–D) is 7–8 days post-eclosion. Genotypes: (A) +/+; +/+; +/+ (OregonR), (B) +/Y; +/+; +/+ (OregonR), (C) w / Y ; chinmo ST /chinmo ST ; +/+, and (D) w/Y; tj-Gal4/+; UAS-chinmo-RNAi/UAS-Dcr-2 (labeled “ tj>chinmo-RNAi ”)

Article Snippet: The following primary antibodies were used: goat anti-Vasa (1:50, dC-13, Santa Cruz), rabbit anti-Vasa (1:3000; gift of Ruth Lehmann, Whitehead Institute, MA, USA), guinea pig anti-Tj (1:5000; gift of Dorothea Godt, University of Toronto, ON, Canada), mouse anti-Fasciclin (Fas) 3 (1:50; Developmental Studies Hybridoma Bank (DSHB), rat anti-DE-cad (1:25, DSHB), mouse anti-Pyd (clone Pyd2) (1:10) (DSHB); rabbit anti-β H -spectrin (1:250) (gift of Claire Thomas, Penn State University, PA, USA); rabbit anti-GFP (1:250) (Invitrogen), mouse anti-β-gal (1:50) (DSHB).

Techniques: Mutagenesis, Labeling