mouse anti histone 2a gamma variant  (Developmental Studies Hybridoma Bank)

Journal: BMB Reports

Article Title: Tau reduction impairs nephrocyte function in Drosophila

doi: 10.5483/BMBRep.2024-0047

Figure Lengend Snippet: Knockdown of dTau disrupts nuclear structure and forms senescence-associated heterochromatin foci (SAHF) in nephrocytes. (A) Nephrocytes were stained with DAPI (blue) to visualize DNA. SAHF, a marker of cellular senescence, was observed in the nephrocytes of dTau RNAi-expressing flies (arrowheads). (B) Quantification of DAPI foci-positive nephrocytes in adult flies. A significant increase in the number of DAPI foci-positive nephrocytes was observed in the dTau RNAi-expressing nephrocytes, compared to the control. Error bars represent the mean ± standard deviation (n = 4 adult flies for each genotype). Statistical significance was determined using Student’s t -test (***P < 0.001). (C) Pericardial nephrocytes (red) in the abdomen of mCD8−RFP-expressing flies under the control of snsGCN−Gal4. Nephrocytes dissected from (3 or 20)-d-old dTau RNAi-expressing flies were stained with anti-histone H2Av antibody (green) and DAPI (blue). dTau knockdown exhibits increased frequencies of γ–H2Av foci in the nephrocytes of 3-d-old flies (arrowheads). Dispersed γ–H2Av foci in 20-d-old dTau RNAi-expressing flies were detected in the cytoplasm of nephrocytes. (D, E) Quantification of nuclear and cytoplasmic γ–H2Av fluorescent signal in the nephrocytes. Error bars represent the mean ± standard deviation (n ≥ 8 nephrocytes for each genotype). Statistical significance was determined using Student’s t -test (**P < 0.01, ***P < 0.001).

Article Snippet: Subsequently, the samples were incubated with the primary antibodies: rabbit anti-DCP-1 antibody (1:100; Cell Signaling Technology, Cat#: 9578, Danvers, MA, USA), mouse anti-Histone H2Av antibody (1:100; DSHB, Cat#: UNC93-5.2.1, Iowa City, IA, USA), and mouse anti-Lamin B (Dm0) antibody (1:100; DSHB, Cat#: ADL67.10, Iowa City, IA, USA) for 12 h at 4°C.

Techniques: Knockdown, Staining, Marker, Expressing, Control, Standard Deviation