Regulation of Yamanaka factors during H5N1 virus infection in A549 cells and HEK293T cells

Figures & data

Table 1. Primer sequences of full-length gene annealing temperature used in construct plasmids.

Gene symbol	Sequence (5′-3′)	Annealing (°C)	Plasmid
NS1	TACGAATTCATGGATTCCAACACTGTGTC	54	3.1
	TACCTCGAGCTTTGGAGAGAGTGGAGGTC
M1	TACAAGCTTATGAGCCTTCTAACCGAGGTCGAA	58	3.1
	TACCTCGAGCTTGAATCGCTGCATTTGCACTC
NP	CCCGGATCCATGGCGTCTCAGGGCACCAAACGA	56	3.1
	CCCCTCGAGATTGTCATATTCCTCTGCATTGTC
PB2	CCCAAGCTTATGGAGAGAATAAAAGAATTAAG	56	3.1
	CCCCTCGAGATAGATGGCCATCCGAATCCTTTTG
HA	TACAAGCTTATGGAGAAAATAGTGCTTCTTCTT	54	3.1
	TACCTCGAGAATGCAAATTCTGCATTGTAACG
NA	TACAAGCTTGCCACCATGAATCCAAATCAGAAGATAATAAC	54	3.1
	TCACTCGAGCTTGTCAATGGTGAATGGCAAC

Table 2. Primer sequences, amplicon size and annealing temperature used in real-time PCR assays.

Gene symbol	Sequence (5′-3′)	Amplicon (bp)	Annealing (°C)
β-actin	GCGGGAAATCGTGCGTGACATT	232	60
	GATGGAGTTGAAGGTAGTTTCGTG
OCT4	GAGGAGTCCCAGGACATCAA	430	60
	ACACTCGGACCACATCCTTC
MYC	AAGTCCTGCGCCTCGCAA	249	60
	GCTGTGGCCTCCAGCAGA
SOX2	TGGACAGTTACGCGCACAT	215	60
	CGAGTAGGACATGCTGTAGGT
KLF4	ACCTACACAAAGAGTTCCCATC	136	60
	TGTGTTTACGGTAGTGCCTG

Figure 1. A549 cells (A) and HEK293T (B) cells infected with H5N1 influenza virus.

Note: Overnight cell cultures seeded on 12-well plates were inoculated with 0.5 MOI, 1 MOI and 2 MOI H5N1 virus and incubated for 36 h. The mock group was inoculated with sterile allantoic fluid.

Figure 2. Expression of Yamanaka factors in A549 cells and HEK293T cells.

Note: Quantitative data (mean ± SEM) from at least three independent experiments were normalized to β-actin. *p < 0.05; **p < 0.01; ***p < 0.001.

Figure 3. Gene expression of Yamanaka factors in A549 cells (A) and HEK293T cells (B) with H5N1 virus infection.

Note: Overnight cell cultures seeded 12-well plates were inoculated with 0.5 MOI, 1 MOI and 2MOI H5N1 virus and incubated for 36 h. Quantitative data (mean ± SEM) from at least three independent experiments were normalized to β-actin. *p < 0.05; **p < 0.01; ***p < 0.001. The mock group was inoculated with sterile allantoic fluid.

Figure 4. Effects of influenza virus proteins NP and PB2 on the gene expression of Yamanaka factors in A549 cells (A) and HEK293T cells (B). Quantitative data (mean ± SEM) from at least three independent experiments were normalized to β-actin. *p < 0.05; **p < 0.01. The mock group was inoculated with sterile allantoic fluid.

Figure 5. Effects of influenza virus proteins NS1 and M1 on the gene expression of Yamanaka factors in A549 cells (A) and HEK293T cells (B).

Note: Differences are not statistically significant.

Figure 6. Effects of influenza virus proteins HA and NA on the gene expression of Yamanaka factors in HEK293T cells.

Note: Statistically non-significant differences following over-expression of HA and NA of influenza virus in HEK293T cells.