Goat anti hpiv3

For immunofluorescence, HeLa cells were cultured on coverslips in 24 well plates overnight and then transfected with N, P, VIM and VIM mutant plasmids. HPIV3 and HPIV3_HA-P were added before transfected. Cells were harvested at the indicated times. 4% paraformaldehyde in 1×PBS was used to fix cells and 0.2% Triton X-100 were added for 20min at room temperature. After being blocked with 3% bovine serum albumin (BSA), primary antibodies diluted in 1% BSA were added and then secondary antibodies diluted in 1% BSA were added at 4°C.The primary antibodies used here were as follows: goat anti-HPIV3 (Abcam), rabbit anti-Flag tag (CST), rat APC-anti-Flag (Biolegend), mouse anti-c-Myc tag (MBL), mouse anti-HA tag (sigma), rabbit anti-HA tag (CST).The secondary antibodies used here were as follows: Alexa Fluor 488 donkey anti-rabbit IgG (Invitrogen), Alexa Fluor 594 donkey anti-mouse IgG (Invitrogen), Alexa Fluor 647 donkey anti-goat IgG (Invitrogen).Image J software were used to assay the fluorescence intensity of the cells in drawing boxes and intensity values were used. In order to visualizing the IBs in cells, HPIV3_HA-P virus was used and the location of HA-P was considered as viral IBs. HA-P and Myc-N co-expression also formed viral IBs in cells without HPIV3 infection. To counting the numbers of large, medium and small viral IBs, almost 30 cells in each group with IBs were counted.