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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.

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