Fig. 3

Cellular co-localization and quantification of NP with M1 and M165A mutant using immunofluorescence confocal microscopy. a/ Co-cultured 293 T and MDCK cells were transfected with eight plasmids for reverse genetic system. After 24 h, the cells were fixed and M1 and M165A were detected using a primary anti-M1 antibody and a secondary antibody coupled to Alexa488 (in green) and NP protein was labelled with anti- NP antibody and a secondary antibody coupled with Alexa555 (in red). Nuclei were stained with DAPI (in blue). NC represents non transfected cells. The intracellular distribution of M1 and NP was imaged by confocal laser scanning fluorescence microscopy (LSM Zeiss 510 Meta). Insets show higher magnification views of the selected areas. The results are representative of three independent experiments. b/ The images obtained from fluorescent microscopy were quantified using Fiji/ImageJ software. The data were delivered from the measurement of 30 cells. The graph indicated mean ± SD. Statistical analysis was performed using t-test. A p-value < 0.05 was considered statistically significant. NP(M1) is NP in the cells transfected with M1, NP(M165) is NP in the cells transfected with M165A mutant