36 though weakly but not no. Number S3: QMEAN scores of the model constructions. QMEAN4 scores of the chimeras (A) and their themes (B) were determined on a Web site (SWISS-MODEL). The amino acid sequence identity between each chimera and its template was also offered in (A). Image_3.TIF (490K) GUID:?29EDA24D-4541-46CB-B850-022038E4B5F2 FIGURE S4: The F stalk domain modifies the HN protein specificity. (A) Effectiveness of fusion induction by chimeric F proteins. The fusion indices given by the chimeric F proteins demonstrated in Number ?Number2D2D are normalized to their cell surface-localization levels. The statistical significance was evaluated by one-way ANOVA as explained in the section Materials and Methods (???< 0.01, = 10). ns, not significant. (B) HN protein specificity of the F proteins. For each chimeric F protein used in Number ?Number2D2D, the fusion index given with CH5-41 was normalized to that given with the SV41 HN protein. The statistical significance was evaluated by one-way ANOVA as ZM 336372 explained in the section Materials and Methods (???< 0.01, = 10). ns, not significant; NA, not applicable. Image_4.TIF (497K) GUID:?6639AD11-9539-4BBD-AAAC-D2DD9F284A5F FIGURE S5: No. 37 is definitely a more SV41 ZM 336372 F-like protein than no. 36. (A) Detection of the F Rabbit polyclonal to AKR1A1 proteins in the plasmid-transfected cells. (Upper panel) long-exposed image of the data presented in Number ?Number3A3A; the cleaved form (F1+2) and unclevaed form (F0) are considered to comigrate with each other under nonreducing conditions. (Lower panel) under reducing conditions, the cleaved form (F1) nearly comigrates with an unidentified cellular protein (ca. 50 kDa), which have been precipitated from the ant-PIV5 F2 polyclonal antibody. In all likelihood, this 50 kDa-protein migrates much slower than F0/F1+2 under non-reducing conditions (A) due to disulfide-mediated association with additional unidentified cellular protein(s). In the parenthesis is definitely indicated the expected position of the uncleaved form, F0. (B) Effectiveness of fusion induction from the F proteins. The fusion indices given by the chimeric F proteins demonstrated in Number ?Number3B3B are normalized to their cell surface-localization levels. The statistical significance was evaluated by one-way ANOVA as explained in the section Materials and Methods (???< 0.01, = 10). Image_5.tif (1.1M) GUID:?FA106F5C-37C2-4930-8E6C-A70FDB9F8C3C FIGURE S6: Immunofluorescent staining the chimeric HN proteins used in Figure ?Figure4B4B. Subconfluent BHK cells produced on glass coverslips in six-well tradition plates were transfected with 2.0 g/well of the pcDLSRa expression vector encoding each HN protein. After 24 h of incubation at 37C, the cells were fixed with 4% paraformaldehyde in PBS, washed three times with PBS, and permeabilized or not permeabilized with 0.1% Triton X-100 in PBS. The HN proteins were visualized by indirect immunofluorescent staining as explained in the section Materials and Methods by using MAb 173-1A or MAb 127A-1. Pub, 100 m. Image_6.TIF (3.9M) GUID:?3A3D9281-CBA2-45B4-A3E9-79777316CF52 FIGURE ZM 336372 S7: Immunofluorescent staining the chimeric HN proteins used in Number ?Figure5A5A. Subconfluent BHK cells produced on glass coverslips in six-well tradition plates were transfected with 2.0 g/well of the pcDLSRa expression vector encoding each HN protein. After 24 h of incubation at 37C, the cells were fixed with 4% paraformaldehyde and the HN proteins were visualized as explained in the story for Supplementary Number S6 by using MAb 173-1A or MAb 127A-1. Pub, 100 m. Image_7.TIF (3.1M).