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Fig. 7 | Virology Journal

Fig. 7

From: Entry of Challenge Virus Standard (CVS) -11 into N2a cells via a clathrin-mediated, cholesterol-, dynamin-, pH-dependent endocytic pathway

Fig. 7

Effect of NH4Cl on CVS-11 infection in N2a cells. a Quantification of cytotoxic effects of NH4Cl on N2a cells ranging from 0 to 80 mM was examined by MTT assay. b N2a cells were pretreated with increasing concentrations (0 mM, 2.5 mM, 5 mM, 10 mM, 20 mM) of NH4Cl for 1 h at 37 °C and infected with CVS-11 (MOI 0.1). At 3 h and 24 h p.i., infected cells were lysed to determine RABV N RNA copy numbers by RT-qPCR. c The cells were pretreated with increasing concentration (0 mM, 2.5 mM, 5 mM, 10 mM, 20 mM) of NH4Cl for 1 h at 37 °C and infected with CVS-11 (MOI 0.1). The cells were lysed and processed for western blot analysis of RABV N protein. GAPDH was used as a loading control. d Relative protein levels were analyzed by using ImageJ. The results are presented as the mean ± SD of three independent experiments. e N2a cells were treated with 20 mM NH4Cl for 1 h and infected with CVS-11 (MOI 0.1). At 24 h p.i., cells were fixed and stained with an FITC-anti-Rabies Monoclonal antibody. Cytoplasm was stained with Evans Blue. Scale bars, 70 μm. f The number of infected cells was counted and percentage of infected cells after drug treated compared to control group was assessed. Five fields of about 200 cells were counted. Means and S.D. values are shown. Statistical significances of the differences are indicated. Student’s t test, p < 0.05 (*); p < 0.01 (**); p < 0.001 (***)

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