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Figure 7 | Virology Journal

Figure 7

From: Influenza A/Hong Kong/156/1997(H5N1) virus NS1 gene mutations F103L and M106I both increase IFN antagonism, virulence and cytoplasmic localization but differ in binding to RIG-I and CPSF30

Figure 7

Host gene expression is differentially affected by 103L and 106I mutation in human and mouse cells. Mouse M1 cells and human A459 cells were wither mock infected with PBS of infected with rHK1-wt, or NS1 F103L, M106I, M106V and F103L + M106I mutants at MOI = 2 in triplicate with extraction of total RNA at 8 hpi. Genes that were significantly increased or decreased 2 fold (≥ 21 or ≤ 2-1) by Affymetrix microarray analysis relative to mock infected cells for any individual virus were analyzed among the viruses. Host genes are represented in heat maps with red and blue according to with the indicated scales for the log 2 values (panels a, b and c). a) Effect of virus infection on gene regulation in mouse M1 cells showing hierarchical cluster analysis of dys regulated genes as well as hierarchical clustering among viruses. b) Effect of virus infection on gene regulation in human A549 cells showing hierarchical clustering of genes and viruses as represented in panel a. Relative to HK-wt the 242 dysregulated genes were significantly upregulated for M106I, M106V and F103L + M106I (** p ≤ 0.01, *** p ≤ 0.001 by t-test) but not F103L (not significantly different (nsd)). c) Transcriptional abundance analysis of genes that are common between human and mouse cells were analyzed as described in panel a. d) Bar graph of up (red bars) and down (blue bars) regulated gene transcripts detected by microarray analysis relative to mock infected M1 and A459 cells infected with the indicated viruses. The number of significantly up and down regulated genes relative to mock infected A549 cells was significantly different for M106I, M106V and F103L + M106I (*p ≤ 0.05 by Chi-squared test), versus HK-wt (reference, r) but not F103L that was not significantly different.

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