Figure 3
Figure 3. Expression of the EBV lytic protein, Zta, contributed to induction of IL-13. (A) BJAB cells were transfected with plasmids expressing EBNA1, EBNA2, LMP1, LMP2A, Zta, or Rta. RNA was extracted from each transfectant at 48 hours after transfection, and IL-13 transcripts were measured by RT-Q-PCR. (B) BJAB cells were infected with pSIN-vector or pSIN-Zta at the MOI indicated. RNA and cell lysates were harvested from each transfectant at 5 days after infection. IL-13 transcripts were measured by RT-Q-PCR (top panel), and Zta expression was detected by Western blot analysis (bottom panel; vertical lines have been inserted to indicate a repositioned gel lane). (C,D) L428 cells were infected with pSIN-Zta or pSIN-vector at an MOI of 4. Cell lysates, RNA, and supernatants were collected on days 4, 5, and 6 after infection. IL-13 transcripts were measured by RT-Q-PCR, and then relative expression of IL-13 transcripts was normalized to the amounts of IL-13 expressed in pSIN-vector transfected cells (panel C top). Zta expression was detected by Western blot analysis, and β-actin was used as an internal control (panel C bottom). Meanwhile, IL-13 protein in supernatants was quantified by ELISA (D). (E) In total, 6 LCLs cells were infected with pSIN-Zta or pSIN-vector lentivirus at an MOI of 4, and RNA and cell lysates were harvested after 7 days. Expression of IL-13 transcripts was detected by RT-Q-PCR, and the relative fold was normalized with the amounts of IL-13 mRNA in pSIN-vector lentivirus control cells (top panel). EBNA2 and Zta expression were measured by Western blot analyses, and β-actin was used as an internal control (bottom panel; vertical lines have been inserted to indicate a repositioned gel lane). (F) LCLs were infected with pLKO-siZta or pLKO-siLuciferase lentivirus, and then RNA and cell lysates were harvested on day 14 after infection. Expression of IL-13 transcripts was detected by RT-Q-PCR, and the relative fold was normalized with the amounts of IL-13 mRNA in pLKO-siLuciferase lentivirus infected cells (top panel). Zta expression was measured by Western blot analysis, and α-tubulin was used as an internal control (bottom panel).

Expression of the EBV lytic protein, Zta, contributed to induction of IL-13. (A) BJAB cells were transfected with plasmids expressing EBNA1, EBNA2, LMP1, LMP2A, Zta, or Rta. RNA was extracted from each transfectant at 48 hours after transfection, and IL-13 transcripts were measured by RT-Q-PCR. (B) BJAB cells were infected with pSIN-vector or pSIN-Zta at the MOI indicated. RNA and cell lysates were harvested from each transfectant at 5 days after infection. IL-13 transcripts were measured by RT-Q-PCR (top panel), and Zta expression was detected by Western blot analysis (bottom panel; vertical lines have been inserted to indicate a repositioned gel lane). (C,D) L428 cells were infected with pSIN-Zta or pSIN-vector at an MOI of 4. Cell lysates, RNA, and supernatants were collected on days 4, 5, and 6 after infection. IL-13 transcripts were measured by RT-Q-PCR, and then relative expression of IL-13 transcripts was normalized to the amounts of IL-13 expressed in pSIN-vector transfected cells (panel C top). Zta expression was detected by Western blot analysis, and β-actin was used as an internal control (panel C bottom). Meanwhile, IL-13 protein in supernatants was quantified by ELISA (D). (E) In total, 6 LCLs cells were infected with pSIN-Zta or pSIN-vector lentivirus at an MOI of 4, and RNA and cell lysates were harvested after 7 days. Expression of IL-13 transcripts was detected by RT-Q-PCR, and the relative fold was normalized with the amounts of IL-13 mRNA in pSIN-vector lentivirus control cells (top panel). EBNA2 and Zta expression were measured by Western blot analyses, and β-actin was used as an internal control (bottom panel; vertical lines have been inserted to indicate a repositioned gel lane). (F) LCLs were infected with pLKO-siZta or pLKO-siLuciferase lentivirus, and then RNA and cell lysates were harvested on day 14 after infection. Expression of IL-13 transcripts was detected by RT-Q-PCR, and the relative fold was normalized with the amounts of IL-13 mRNA in pLKO-siLuciferase lentivirus infected cells (top panel). Zta expression was measured by Western blot analysis, and α-tubulin was used as an internal control (bottom panel).

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