Afro Samurai avenged his father and found a life of peace. But the legendary master is forced back into the game by a beautiful and deadly woman from his past. The sparks of violence dropped along Afro’s bloody path now burn out of control – and nowhere are the flames of hatred more intense than in the eyes of Sio.
| Actor | Role |
|---|---|
 Samuel L. JacksonAfro / Ninja-Ninja (voice) | Afro / Ninja-Ninja (voice) |
 Lucy LiuSio (voice) | Sio (voice) |
 Yuri LowenthalJinno (voice) | Jinno (voice) |
 Mark HamillBin / Oden Shop Master (voice) | Bin / Oden Shop Master (voice) |
 Jeff BennettBrother 3 (voice) | Brother 3 (voice) |
 S. Scott BullockProfessor Dharman / Kidnapper (voice) | Professor Dharman / Kidnapper (voice) |
 Grey DeLisleTomoe / Ogin (voice) | Tomoe / Ogin (voice) |
 Greg EaglesAfro's Father (voice) | Afro's Father (voice) |
 Zachary GordonKotaro (voice) | Kotaro (voice) |
 Liam O'BrienShichogoro (voice) | Shichogoro (voice) |
 RZADJ (voice) | DJ (voice) |
 Phil LaMarrYoung Afro (voice) | Young Afro (voice) |
 Ariel WinterYoung Sio (voice) | Young Sio (voice) |
 Kevin Michael RichardsonBlacksmith / Takimoto (voice) | Blacksmith / Takimoto (voice) |
 Steve BlumMan A / Assassins (voice) | Man A / Assassins (voice) |
 Dave WittenbergAdolescent / Fencing Brother (voice) | Adolescent / Fencing Brother (voice) |