Skip to main content
. 2017 Apr 13;12(4):e0175009. doi: 10.1371/journal.pone.0175009

Fig 4. MFN2 siRNA attenuates the protective effects of RSV against the apoptosis induced by CSE in HBE cells.

Fig 4

(A) The transfection efficiency of HBE cells was detected by fluorescence microscopy, which showed that more than 90% of cells were labeled (red fluorescence). (B-C) Real-time qPCR and western blot analysis were used to test three MFN2 RNA interference target sequences for gene silencing efficiency. The results show that the first silence sequence (si-h-mfn2_001: CGGCAAGACCGACTGAAAT) exhibited the highest efficiency, and the optimal intervention time was 72 h. Therefore, the first RNA interference target sequence was selected for the following experiments. (D) CSE-induced apoptosis was markedly attenuated by RSV pretreatment (RNAi + RSV + CSE); however, this protective effect was abolished by MFN2 gene knockdown (MFN2 siRNAi+ RSV + CSE). (E) Western blot analysis showed apoptosis marker molecules in all of the groups. BAX began to translocate to mitochondria in the CSE group, and cytochrome C in the cytoplasm began to increase in the CSE group compared with that in the MFN2 siRNA group (p<0.05). However, the effects of RSV to prevent the release of the cytochrome C from the mitochondria to the cytosol and prevent the translocation of the BAX from the cytosol to the mitochondria were decreased. There were no differences between the MFN2 siRNA+ CSE group and the MFN2 siRNA+ RSV+CSE group (p>0.05). (F) Western blot analysis showed that the RSV pretreatment did not increase the MFN2 levels in the MFN2 siRNA groups compared with the negative control siRNA groups. *: p<0.05, **: p<0.01, ***; p<0.001, #: p>0.05.