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The first RAF base to receive Mustang IIIs was at Gravesend in Kent. The Mustang III initially equipped No. 65 Squadron in late December of 1943, followed by No. 19 Squadron in March of 1944. Later the Mk. III also equipped Nos 64, 65, 66, 93, 94, 112, 118, 122, 126, 129, 165, 234, 237, 241 249, 250, 260, 268, 306, 309, 315, 316, 345, 430, 441, 442, and 516 Squadrons and No. 541 Squadron of RAF Coastal Command.

The RAF Mustang IIIs began operations late in February 1944, escorting US heavy bombers as well as both US and RAF medium bombers.