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Playing in the Championship, Southampton stunned Arsenal despite the London side dominating possession and creating numerous chances.
Southampton took the lead in the 35th minute through Ross Stewart, leaving Arsenal trailing at the break despite their control of the match.
Substitute Viktor Jukkasas brought Arsenal level in the 68th minute, but the Gunners failed to capitalize on their dominance.
Chay Charles struck in the 85th minute to secure a memorable win for Southampton, handing Arsenal only their fifth defeat of the season ahead of their Champions League clash with Sporting.