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Level up Robin, Franky & Brook to level 3 in Story mode to unlock their pre-timeskip's second support attack. Robin's second attack is 'Wing', Franky's is 'Ballad' and Brook's is 'Yahazugiri'.
When you have completed the 'Thousand Arena Battle Royale' and 'One To One' challenges under the Shin Sekai difficulty setting 5 times each with different characters the '1 vs. 100' challenge becomes available. The '1 vs. 100' challenge consists of a series of 100 short fights where you participate alone and cannot recover between each fight. After every 10 victories you will get a piece of meat.
This very hard difficulty setting for the two main tournaments of the 'Thousand Arena mode' ('Battle Royale' & 'One To One') becomes available when you finish the tournament in Grand Line difficulty (Hard mode) with 5 different characters.
The following characters are not unlocked through Story mode but by completing the indicated task.
Unlock Soldier Docking Emperor:
Complete 'Thousand Arena Battle Royale' in any difficulty with 45 characters.
Unlock Chopperman:
Complete 'Thousand Arena One To One' in any difficulty setting with 45 characters.
Unlock Neptune:
Connect to Nintendo WFC and select 'Download Content' to unlock a secret Story Mode level. Then complete the level to unlock Neptune.
Unlock Shirahoshi:
Connect to Nintendo WFC and select 'Download Content' to unlock a secret Story Mode level. Then complete the level to unlock Shirahoshi.
Unlock Main Character Support Versions:
Either execute ALL character moves in Training mode or complete a 'Thousand Arena' tournament with the character you want.