(PDF 146 KB) Additional file 10: Figure S7: Schematic diagram of the Rad3 helicase family in G. lamblia. The representation is to scale. Inset: sequence LOGO view of the consensus amino acids. The height of each amino acid represents the degree of conservation. Colors indicate properties of the amino acids, as follows: green (polar), blue (basic), red (acidic)
and black (hydrophobic). (PDF 148 KB) Additional file 11: Figure S8: Western blot of GDC-0994 purchase trophozoites grown under proliferating conditions and after induction to encyst. Total protein extracts from trophozoites grown under normal proliferating conditions (Normal) or after 16hs induction in encystation medium (Encyst) were separated using a 10% SDS-polyacrylamide gel and MI-503 order transferred to a PVDF membrane. The membrane was incubated with a monoclonal antibody against CWP2. The iqual loading of the samples is shown in the figure at the right with a Ponceau S staining. The numbers indicate the molecular weight of protein standards in kDa. (PDF 97 KB) Additional file 12: Figure S9: SAGE (Serial Analysis of Gene Expression) data. The VRT752271 graph represents the sense tag
percentage from Giardia trophozoites (white bar) and four different encystation times (4, 12, 21 and 42 hours; grayscale bars). Under each ORF it is indicated if these ORFs were up-regulated (green up arrow), down-regulated (red down arrow) or remained unmodified (equal sign). A line graph is also provided for a better identification of the expression pattern. The colored boxes
represent our RT-qPCR results (with the same color code), divided into families. The asterisk under each box stands for a correlation between the SAGE and the RT-qPCR data. (PDF 236 KB) Additional file 13: Figure S10: Western blot during antigenic variation induction. Trophozoites were incubated for the indicated times with a 1:10.000 dilution of mAb 5C1directed against VSP-1267, mAb 7D2 against Cyst Wall Protein 2 or without antibody (Control). Total protein was electrophoresed, transferred to a PVDF membrane and incubated with a mAb against the VSP-1267. The molecular Protirelin weights of standards are indicated in kDa. (PDF 71 KB) Additional file 14: Table S4: Accession numbers. The table indicates a complete list of proteins cited in the manuscript, the organism it is derived and the NCBI Reference Sequence Number. (XLSX 10 KB) References 1. Abdelhaleem M: Helicases: an overview. Methods Mol Biol 2010, 587:1–12.PubMedCrossRef 2. Linder P, Jankowsky E: From unwinding to clamping – the DEAD box RNA helicase family. Nat Rev Mol Cell Biol 2011, 12:505–516.PubMedCrossRef 3. Singleton MR, Dillingham MS, Wigley DB: Structure and mechanism of helicases and nucleic acid translocases. Annu Rev Biochem 2007, 76:23–50.PubMedCrossRef 4. Kainov DE, Tuma R, Mancini EJ: Hexameric molecular motors: P4 packaging ATPase unravels the mechanism. Cell Mol Life Sci 2006, 63:1095–1105.PubMedCrossRef 5.