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. 2014 Aug 23;11:148. doi: 10.1186/s12974-014-0148-9

Figure 4.

Figure 4

Downregulation of CNPase after CNPase siRNA transfection in BV-2 cells. (A) There was no noticeable change in external morphology in BV-2 cells when transfected with either control small interfering RNA (siRNA) or CNPase siRNA, and when compared with the non-transfected control cells under the phase-contrast microscope. (B) The viability of BV-2 cells transfected with control siRNA and CNPase siRNA is 92% and 91%, respectively, against the non-transfected control value. (C) Reverse transcription polymerase chain reaction analysis shows that the efficiency of siRNA (160 #)-mediated suppression of CNPase is about 84% while that of siRNA (161 #) is about 94% compared to negative control (normalized with β-actin). (D) The upper panel shows the specific Western band of CNPase and β-actin proteins. The lower panel shows bar graphs depicting significant changes in the optical density of different groups. Note the remaining CNPase protein expression in CNPase siRNA (161 #) transfected BV-2 cells is about 25% compared to the control siRNA transfected BV-2 cells. (E) Immunofluorescence images show CNPase immunoreactivity is markedly reduced in CNPase siRNA transfected BV-2 cells compared to negative control. *P < 0.05, **P < 0.01 and ***P < 0.001. The values represent the mean ± SD in triplicate. Scale bars = 100 μm (A) and 20 μm (E). CNPase, 2′,3′-cyclic nucleotide 3′-phosphodiesterase; DAPI, 4′,6- diamidino-2-phenylindole.