anti hemagglutinin  (Cell Signaling Technology Inc)

Journal: Poultry Science

Article Title: eIF3m promotes fowl adenovirus serotype 4 replication via interacting with ORF1B protein

doi: 10.1016/j.psj.2026.106566

Figure Lengend Snippet: eIF3m interacted with FAdV-4 ORF1B protein in LMH cells. (A and B) eIF3m interacted with exogenous ORF1B. LMH cells co-transfected with 1 μg/well of p3 × flag-ORF1B and 1 μg/well pCAGGS-HA-eIF3m were lysed at 48 h, and cell lysates were immunoprecipitated with an anti-flag (A) or anti-HA (B) antibody followed by western blot with anti-HA and anti-flag antibodies. (C) The interaction of eIF3m with endogenous ORF1B. LMH cells transfected with 1 μg/well of pCAGGS-HA-eIF3m or empty vector were infected with CH/HNJZ/2015 at an MOI of 0.01 at 24 hpt. Cell lysates were immunoprecipitated with an anti-HA antibody followed by western blot with anti-HA and anti-ORF1B antibodies. (D) eIF3m and ORF1B were co-localized in cytoplasm. LMH cell co-transfected with 1 μg/well of pEGFP-ORF1B and 1 μg/well of pDsRed-eIF3m were fixed at 24 h. After stained with DAPI, the fluorescence was observed by using a confocal microscope.

Article Snippet: Commercial antibodies used in this study included rabbit anti-flag monoclonal antibody (240568AA1, proteintech), mouse anti-HA monoclonal antibody (66006-2-Ig, proteintech), rabbit anti-GAPDH monoclonal antibody (A19056, ABclonal).

Techniques: Transfection, Immunoprecipitation, Western Blot, Plasmid Preparation, Infection, Staining, Fluorescence, Microscopy

Journal: eLife

Article Title: Genetic and physical interactions reveal overlapping and distinct contributions to meiotic double-strand break formation in C. elegans

doi: 10.7554/eLife.96458

Figure Lengend Snippet: ( A ) Western blot showing specificity of the XND-1 antibody. Western analysis was performed on protein extracts from wild-type N2 worms and two independent xnd-1 mutant strains. A distinct band corresponding to XND-1 (red square) is detected only in the N2 extract, confirming the specificity of the antibody. No signal is observed in the mutant strains, consistent with loss of xnd-1 expression (see also ). ( B ) 3xHA::HIM-17 and anti-XND-1 staining do not overlap with one another or with the DNA axes. Shown here are 3D renderings of confocal stacks from the mitotic zone, early-middle pachytene, and mid-late pachytene regions. ( C ) Localization of XND-1 is normal in him-17(ok424 M-Z- ) mutants (anti-XND-1, pink; DNA/DAPI, green) (top). Localization of 3xHA::HIM-17 is unaffected in xnd-1(ok709 ) mutants (anti-HA, pink; DNA/DAPI, green) (bottom).

Article Snippet: Mouse monoclonal anti-HA (Cell Signaling), chicken polyclonal anti-GFP (AbCam), mouse monoclonal anti-FLAG HRP-conjugated (Sigma), and polyclonal anti-XND-1 ( ) antibodies were diluted in blocking solution at 1:1000, 1:5000, 1:2000, and 1:2500, respectively, and left to incubate overnight at 4°C.

Techniques: Western Blot, Mutagenesis, Expressing, Staining

Journal: eLife

Article Title: Genetic and physical interactions reveal overlapping and distinct contributions to meiotic double-strand break formation in C. elegans

doi: 10.7554/eLife.96458

Figure Lengend Snippet: ( A ) Immunofluorescence analysis of HIM-5::HA in dsb-1 mutants. DAPI (blue) stains DNA, anti-HA marks endogenously tagged HIM-5 (green), and pHTP-3 S285 (red) labels chromosome axes. HIM-5 appears localized in nuclei in pre-meiotic stages (indicated by arrows). However, after the transition zone (TZ), HIM-5 loses its nuclear localization. ( B ) Immunofluorescence analysis of HIM-5::HA in him-5::3XHA control . Top: DAPI (blue) stains DNA, anti-HA marks endogenously tagged HIM-5 (green), and pHTP-3 S285 (red) labels chromosome axes. Bottom: Zoomed-in region where HIM-5 (yellow) can be observed localizing in pre-TZ nuclei through mid-pachytene. ( C ) Top: C. elegans gonad fixed and stained with DAPI to show the organization and distribution of the nuclei along the Prophase I. Bottom: Live imaging of nuclei in the transition zone (leptotene–zygotene) and middle-pachytene. eaIs15 (Ppie-1::him-5::GFP) is visualized in freshly dissected gonads by GFP fluorescence (green), and DNA by DRAQ5 (red). In dsb-1 mutants, HIM-5 is nuclear in the transition zone and then only appears in cytoplasmic puncta by middle pachytene.

Article Snippet: Mouse monoclonal anti-HA (Cell Signaling), chicken polyclonal anti-GFP (AbCam), mouse monoclonal anti-FLAG HRP-conjugated (Sigma), and polyclonal anti-XND-1 ( ) antibodies were diluted in blocking solution at 1:1000, 1:5000, 1:2000, and 1:2500, respectively, and left to incubate overnight at 4°C.

Techniques: Immunofluorescence, Control, Staining, Imaging, Fluorescence