Figure 4
Induction of ISG by HIV Tat SF2 in K562 and U5A cells (4E-4H) cells. Detection of Tat RNA expression by RT-qPCR in K562 and U5A cells 16 and 30 h (A) and 30 h (E) after adenoviral infection. B. Detection of ISG RNA expression by RT-qPCR after adenoviral infection in K562-DC at 16 and 30 h. Statistical significance was detected by T Test when comparing TatSF2 and TatSF21-58 values at 16 h for IRF-7 (p = 0.02), and for the ISG RNA values grouped together (p = 0.0007); at 30 h for IP-10 (p = 0.04), MCP2 (p = 0.05), MIG (p = 0.049), TRAIL (p = 0.01), and for the ISG RNA values grouped together (p = 2.19E-06). C. IP10 ELISA of TatSF2 and TatSF21-58 expressing K562 supernatants; p = 0.0002 by ANOVA and Bonferroni’s multiple comparison test was significant for all pairs except UI and tTA; significant differences were also found by T-Test between, tTA and TatSF2 (p = 0.05), tTA and TatSF21-58 (p = 0.0001) and TatSF2 and TatSF21-58 (p = 0.047). D. Protein analysis of TRAIL levels by ICS in K562: tTA and TatSF2 (p = 0.043, T-Test) and TatSF2 and TatSF21-58 (p = 0.041). F. Detection of ISG RNA expression by RT-qPCR after adenoviral infection in U5A cells at 30 h. Statistical significance was detected when comparing TatSF2 and TatSF21-58 values for IRF-7 (p = 0.03,T-Test), IP-10 (p = 0.01), TRAIL (p = 0.02), and for the cumulative values of all the ISG (p = 0.0046). G. IP10 ELISA of TatSF2 and TatSF21-58 of infected U5A supernatants; p = 0.0008 by 1-way ANOVA, Bonferroni’s multiple comparison test was significant for all pairs except UI and tTA; significant differences were also found by T-Test between tTA and TatSF21-58, (p = 0.003), tTA and TatSF2 (p = 0.008) and TatSF2 and TatSF21-58 (p = 0.01). H. Protein analysis of TRAIL levels by ICS in K562: tTA and TatSF2 (p = 0.03, T-Test) and TatSF2 and TatSF21-58 (p = 0.04, T-test). RNA fold induction represents the ratios between the RT-PCR result obtained for cells expressing Tat or Tat1-58 and that obtained for Ad-tTA infected cells. Ct values in all samples were normalized using β-actin expression levels. Tat amount in the 30 h wild-type TatSF2 infected sample was assigned an arbitrary value of 1.