Symmetric Tree, ClustalW. Divergence x 0.5 Divergence x 1 Divergence x 2. Alignment length

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1 ONLINE APPENDIX Talavera, G., and Castresana, J. (). Improvement of phylogenies after removing divergent and ambiguously aligned blocks from protein sequence alignments. Systematic Biology, -. Symmetric Tree, ClustalW Divergence x. Divergence x Divergence x ClustalW, Gblocks relaxed ClustalW, Gblocks stringent Online Appendix - Figure. Average Robinson-Foulds distances to the real from the calculated with ClustalW alignments (solid, red line), and the same alignments after treatment with Gblocks relaxed (dotted, blue line with diamonds) and stringent (dotted, green line with square symbols) conditions. The symmetric with three different divergence levels was used for the simulations. Trees were reconstructed by ML, NJ and parsimony.

2 Intermediate Tree, ClustalW Divergence x. Divergence x Divergence x ClustalW, Gblocks relaxed ClustalW, Gblocks stringent Online Appendix - Figure. Average Robinson-Foulds distances to the real from the calculated with ClustalW alignments (solid, red line), and the same alignments after treatment with Gblocks relaxed (dotted, blue line with diamonds) and stringent (dotted, green line with square symbols) conditions. The intermediate with three different divergence levels was used for the simulations. Not all alignment lengths were simulated. Trees were reconstructed by ML, NJ and parsimony.

3 Symmetric Tree, Mafft Divergence x. Divergence x Divergence x Online Appendix - Figure. Average Robinson-Foulds distances to the real from the calculated with Mafft alignments (solid, red line), and the same alignments after treatment with Gblocks relaxed (dotted, blue line with diamonds) and stringent (dotted, green line with square symbols) conditions. The symmetric with three different divergence levels was used for the simulations. Trees were reconstructed by ML, NJ and parsimony.

4 Intermediate Tree, Mafft Divergence x. Divergence x Divergence x Online Appendix - Figure. Average Robinson-Foulds distances to the real from the calculated with Mafft alignments (solid, red line), and the same alignments after treatment with Gblocks relaxed (dotted, blue line with diamonds) and stringent (dotted, green line with square symbols) conditions. The intermediate with three different divergence levels was used for the simulations. Not all alignment lengths were simulated. Trees were reconstructed by ML, NJ and parsimony.

5 KOG Divergence x. Divergence x Divergence x KOG Online Appendix - Figure. Average Robinson-Foulds distances to the real from the ML calculated with Mafft complete alignments (solid, red line), and the same alignments after treatment with Gblocks relaxed (dotted, blue line with diamonds) and stringent (dotted, green line with squared symbols) conditions. The asymmetric with three different divergence levels was used for the simulations. Profiles of rate heterogeneity derived from two different proteins, KOG and KOG, were used.

6 amino acids amino acids Asymmetric Intermediate Symmetric Asymmetric Intermediate Symmetric ML Divergence x. Divergence x Divergence x NJ Divergence x. Divergence x Divergence x Divergence x. Divergence x Divergence x ClustalW, GBlocks relaxed ClustalW, GBlocks stringent Mafft, GBlocks relaxed Mafft, GBlocks stringent Online Appendix - Figure. Overall alignment strategies that give rise to the statistically best topologies. The complete alignment is represented by a black rectangle (ClustalW) or oval (Mafft), and the two different Gblocks strategies by rectangles or ovals with stripes that represent the stringency of the selection.

7 Bootstrap analysis Symmetric Tree - NJ A 9 Divergence x. Divergence x Divergence x ClustalW, Gblocks relaxed ClustalW, Gblocks stringent B 9 Online Appendix - Figure. Average bootstrap values of NJ s obtained from ClustalW (A) and Mafft (B) alignments simulated from the symmetric with three different divergence levels. Complete (solid, red line), Gblocks relaxed (dotted, blue line with diamonds) and Gblocks stringent (dotted, green line with squared symbols) alignments are shown.

8 Bootstrap analysis Asymmetric Tree - ML A 9 Divergence x. Divergence x Divergence x ClustalW, Gblocks relaxed ClustalW, Gblocks stringent B 9 Online Appendix - Figure. Average bootstrap values of ML s obtained from ClustalW (A) and Mafft (B) alignments simulated from the asymmetric with three different divergence levels. Complete (solid, red line), Gblocks relaxed (dotted, blue line with diamonds) and Gblocks stringent (dotted, green line with squared symbols) alignments are shown.

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