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. 2017 Dec 13;8(12):3215. doi: 10.1038/s41419-017-0053-0

Fig. 3. Overexpression of MKK7 or its phosphomimetic promotes IBV-induced apoptosis.

Fig. 3

a H1299 cells were transfected with pXJ40-FLAG, pXJ40FLAG-MKK7, pXJ40FLAG-MKK7-KM, pXJ40FLAG-MKK7-3E, or pXJ40FLAG-MKK7-3A. After 24 h, the cells were irradiated with UVC (10 mJ) and incubated for another 1 h. Protein lysates were harvested and were subjected to Western blot analysis using the indicated antibodies. Beta-actin was included as loading control. Sizes of protein ladders in kDa were indicated on the left. Degree of JNK phosphorylation was calculated as in Fig. 1a. The experiment was repeated three times with similar results, and the result of one representative experiment is shown. b H1299 or Huh-7 cells were transfected as in a before being infected with IBV at MOI~2 or being mock infected for 20 h. The cells were harvested and were subjected to Western blot analysis using the indicated antibodies. Beta-actin was included as loading control. Degree of JNK phosphorylation was calculated as in Fig. 1a. Percentage of PARP cleavage [PARP Clv. (%)] was calculated as the intensity of cleaved PARP [PARP(Cl)] divided by the total intensities of full length PARP [PARP(FL)] and PARP(Cl).The experiment was repeated three times with similar results, and the result of one representative experiment is shown. c The culture supernatants of IBV-infected Huh-7 cells in b were subjected to plaque assay analysis. Virus titers were expressed as the logarithm of plaque-forming units (PFU) per ml of supernatants.