International Journal of Molecular Veterinary Research, 2024, Vol.14, No.5, 185-193 http://animalscipublisher.com/index.php/ijmvr 188 Figure 2 Characteristics of ASFV pI73R. I73R is a conserved gene and is transcribed and expressed in the early stage of the ASFV replication cycle (Adopted from Liu et al., 2023) Image caption: (A) Heatmap demonstrating the clustering of transcriptional patterns of the ASFV genome at various time points. The abundance of ASFV transcripts was expressed as fragments per kilobase million (FPKM) and indicated using a color key (blue and red correspond to decreased and increased transcriptional levels, respectively). Each column represents one sample, while each row represents the results of hierarchical clustering. (B) The transcript levels of the top 20 genes during ASFV replication. (C) The transcriptional levels of the I73R gene at various time points. The RNAs encoding the open reading frames of I73RB646L, and CP204L were isolated from BMDMs infected with ASFV-GZ at 3, 6, 9, 12, 15, and 18 h postinfection. (D) The protein expression levels at various time points. BMDMs were infected with ASFV-GZ at an m.o.i of 1 for 0, 3, 6, 9, 12, 15, and 18 h. The expression levels of pI73R, p72 and p30 were detected using anti-pI73R, anti-p72 and anti-p30 antibodies, respectively. β-actin was used as the loading control. (E) Subcellular localization of pI73R in BMDMs during ASFV infection. BMDMs were infected with ASFV-GZ (m.o.i = 1) for 6, 12, and 18 hpi and reacted with anti-pI73R monoclonal antibodies and Alexa 488-conjugated goat anti-mouse IgG secondary antibody (green). The nuclei were stained with DAPI (blue) (63×). Note: White arrows indicate the location of the cellular virus factories (VF) formed after ASFV infection. (F) Subcellular localization of pI73R in HeLa cells. HeLa cells were transfected with recombinant pI73R-Flag plasmid and probed with anti-Flag antibodies. The nucleus were stained with DAPI. Green fluorescent protein (GFP) (pseudocolored in green), DAPI (pseudocolored in blue) images were captured using a confocal microscope (63×) (Adopted from Liu et al., 2023) 4 Molecular Mechanisms Underlying ASFV Virulence 4.1 Immune modulation by ASFV virulence genes African swine fever virus (ASFV) employs various strategies to modulate the host immune response, which is crucial for its virulence. The virus encodes several genes that interfere with host immune signaling pathways. For instance, the A238L gene regulates NFκB and NFAT pathways, which are critical for immune response modulation (Gallardo et al., 2018). Additionally, the MGF360-9L gene antagonizes the JAK/STAT signaling
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