All sorts of stuff.
Minor changes:
1. Fully enclosed wheel wells (tested in the Mk III)
2. Retractable tail wheel (happened in the Mk VIII)
3. Fully flush riveted construction (happened in the Mk VIII)
4. Leading edge fuel tanks (happened in the Mk VIII)
5. Larger forward fuel tanks (happened in the Mk VIII)
6. Fit Hispano Mk V cannon (happened in the Mk 21)
7. Fit inboard .50 cal (happened in the Mk IXe)
Moderately easy changes:
7. Re-angle front windscreen and fit curved 'Speed Spitfire' window fairing (tested on the Mk XIV, showed 8 mph improvement in speed)
8. Fit 29 imp gal rear fuselage tank (happened in the Mk V)
9. Re-design the aircraft to use US-style aircraft fasteners (much lower panel gaps on US fighters)
10. Re-profile the nose skinning to Griffon-engine style 1 piece skin
11. Lengthen the nose profile slightly to add more fuel and aid the CoG issue
Difficult changes:
12. Reprofile the radiators to take better advantage of boundary layer and Meredith effect
13. Partially laminar flow wing
Pretty much bang on. except the wing.
The Spit wing very, very good and had lower drag than the Mustang's in several flight regimes (overall the Spit was better on the climb and worse on the dive, roughly similar on level flight(ie a low Cl), better at high speeds with better mach induced drag (where drag stops being linear with the square of the speed, and become exponential).
Therefore the lower speed for the same power came down to radiator drag and detail drag.
Small details matter. The 20mm cannon Mk Vs were 5mph slower than the Mk Vs with 8x0.303.
The bulletproof flat windscreen cost (with a rear mirror) about 10mph (ditto the Mosquito fighters, fighter/bombers, vs recon and bombers).
It got worse with speed as it had a critical mach limit of only about 0.6 (and a bit from memory), therefore the windscreen drag got worse at high speeds.
Radiator drag. The Spit, like the Me-109 used the first versions of utilising the Meridith effect to reduce radiator drag. The Mustang was a generation ahead.
It's placement was far better, being in the fuselage meant less turbulent air and thus better flow control and cooling efficiency. And the design was better, with variable shuttering vs the simple limited settings for the Spit and 109.
Yes the Mustang still had some net radiator drag, but it was very low.
Even one of the designers of the Mustang discounted the wing (which Supermarine found out when it made its own 'laminar' flow wing) as being the reason for its speed. Wind tunnel tests confirm this.
If you take, on similar powers, a 30mph difference in speed (yes it varied a bit for model to model and configuration to configuration, that's a rough average) then 50% was from radiator efficiency, the other 50% was details. I'd take 25mph as a gap to aim for, because the Mustang would also have lost 5mph with similar cannons.
The prototype Spit Mk III, with better radiators and lots of all those details cleaned up was rated for about 400mph compared to 375 for a similarly powered normal Spit. That's that magic 25mph region again.
Supermarine actually proposed moving the radiators to the same location as in the Mustang, and estimated (for a 60 series engine) 430mph. MAP rejected it due to production disruption issues and then came along the Griffon.
So they could have done it. If RR hadn't been able to keep increasing Merlin power and thing bringing on the Griffon then they would probably have been forced to. But they took the easy way of more power being available as it was easier to re-engine the Spit (a wonderful thing about its design) than do other changes.