Zero post-translational acetyl groups + 2 methyl groups
4
7
3
10 5 6
2
8 9
1
1600
13
11 12
1610
1620
14
1630
15 16
1640
1600
1650
1610
1620
m/z
1630
1640
1650
1600
1610
1630
1640
1650
m/z
m/z
T=0
1620
T=30 min
T=15 min
4 post-translational acetyl + 2 methyl groups
Post-translational acetylations 4 acetyl 3 acetyl
Number of post-translational methyl groups
2 acetyl
0 1 2 0 1 2 0 1 20 1 2 0 1
1 acetyl 0 acetyl
1600
1610
1620
1630
1640
m/z
T=60 min
1650
1600
1610
1620
1630
1640
m/z
T=120 min
1650
1600
1610
1620
1630
1640
m/z
T=180 min
1650
Supplementary Figure 1. Fourier Transform Mass Spectometry of intact acidextracted histone H4 molecules. The serial profiles represent the increase in histone H4 acetylation over time in cultured HCT116 cells in response to high dose HDAC inhibition with 500 nM TSA. At T0 the predominant histone H4 peak is unacetylated, whereas at T180 (minutes) the predominant histone H4 molecule in the spectrum is 4 acetylated (and 2 methylated). The table shows the modifications of all peaks within the spectrum.
Supplementary Figure 2
Ratio of emission at 525 nM vs. 685 nM
120000 110000 100000 90000
Standard curve Control at neutral pH
80000
D-Lactate 30 mM L-lactate 30 mM
70000 60000 50000 40000 6
6.5
7
7.5
pH
8
8.5
Supplementary Figure 2. Internal pH is not affected by a 4 hour exposure to 30 mM L-lactate or 30 mM D-lactate.
Supplementary Figure 3
Probability of deregulation
A Up regulated Series1
0.2 0.18 0.16
Series2 Down
regulated
Up or Series3
down regulated
0.14 0.12 0.1 0.08 0.06 0.04 0.02 0
4550
5055
5560
6065
6570
7075
7580
8085
8590
9095
95100
100200
200500
500103
103 >5x103 5x103
500103
103- >5x103 5x103
B
Probability of deregulation
Expression levels of probes in untreated cells
0.3
0.25
0.2
0.15
0.1
0.05
0 4550
5055
5560
6065
6570
7075
7580
8085
8590
9095
95100
100200
200500
Expression levels of probes in untreated cells
Supplementary Figure 3. – The probability of being significantly deregulated (t-test, P
