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Play as Cow Girl Anne
Successfully complete survival mode two or more times.

Play as Way of the Samurai 1 and 2
Successfully complete all levels (including the extra levels) under the super hard difficulty setting.

Other bonus characters
Find the wanted posters in the following levels to unlock the corresponding character.

Faceless
Hard level 8 (Saloon): Found over the chandelier.

Donna
Hard level 11 (Cactus Gulch): Get to a position where the train station is behind you. Look on the terrace of the building to the left and diagonal from your location.

Killer
Hard extra level 2 (Coal Mine): Above the coal mine entrance.

Nathan
Hard level 9 (Coal mine): From the starting location, search the building directly in front, and beyond the slippery ground.

Jean
Hard level 12 (Wilderness): Found in the air next to the bridge.

Rando
Hard level 15 (Fort): Search the back of the right side of the fort.

Goldberg (normal)
Hard level 16 (Goldberg Residence): Look behind the back stairs inside Goldberg's room.

Goldberg (battle)
Easy extra level 1 (Fort): Found diagonally above the statue.

Harvey
Hard extra level 4 (Coal Mine): Look behind the bridge.

General
Hard level 7 (Cactus Gulch): Search inside the small room along the tracks.

Lanky
Hard level 14 (Wilderness): Look around the entrance to the level.

Oaf
Hard level 2 (Saloon): Search down by the stairs.

Ripper
Hard level 1 (Cactus Gulch): Found above the general store next to the saloon.

Eve
Hard level 3 (Cactus Gulch): Hidden in the back of the small path next to the train station.

Bart
Hard level 6 (Fort): Found in an opening by the entrance to the courtyard.

Vile
Hard level 4 (Ghost Town): From your starting position, look behind the road blocking sign in front of you.

Little Baby
Hard level 5 (Goldberg Residence): Found behind a door.

Faceless (type 2)
Hard level 13 (Saloon): Look on the pipe next to the stage.

Claudia
Hard extra level 4 (Coal Mine): Go past the bridge and enter the room with a crane. It is on the wall to the right.

Anne
Hard level 10 (Ghost Town): Found in mid-air in the cemetery.

Joey
Hard level 9 (Coal Mine): Go past the bridge and look above the dead end to the right.

Ralph
Hard level 16 (Goldberg Residence): On the first floor, search the pipe on the back left.

Urchin
Normal extra level 5 (Wilderness): Look at the top of the hill.

Sweetheart
Normal level 16 (Wilderness): Look in the dining room, behind the screen.

Brat
Hard level 10 (Ghost Town): Search inside the bathroom.