Table 2.

ASK1 phosphorylation dynamics in response to increasing concentration levels of HNE and H₂O₂.

		Peak area ratio HNE1		Peak area ratio H2O21
Module	Peptide Modified Sequence2	0 to 10μM	0 to 50μM	0 to 500μM	0 to 5000μM
	S82 GRGSSVGGGSR			0.7	4.9**
	S82 GSSVGGGSRR			0.7	6.5***
	S82 GSSVGGGSR			0.9	2.4
A	S83 GRGSSVGGGSRR			0.7	3.6****
	S82|S83 GRGSSVGGGSR	0.9	0.7
	S82|S83 GRGSSVGGGSRR	0.6	0.6**
	S82|S83 GSSVGGGSRR	0.9	0.6
B	YT140 LDFGETTVLDR			2.1*	2.5
C	S269 VAQASSSQYFR			1.3	1.7
D	Y355 FHYAFALNR			0.2*	0.4*
	T947|S952 KKTQPKLSALSAGSNEYLR	0.7*	0.1****
E	T947|S952 KKKTQPKLSALSAGSNEYLR	1.0	0.2**
	S952KKKTQPKLSALSAGSNEYLR			1.1	1.9
	S955 LSALSAGSNEYLR			1.5**	2.8***
	S958 LSALSAGSNEYLR			1.6*	2.3**
F	S955|S958 KTQPKLSALSAGSNEYLR	7.0***	1.1
	S955|S958 LSALSAGSNEYLR	0.9	0.7
	S955|S958 TQPKLSALSAGSNEYLR	2.4**	0.7
	T1000|S1004 TRAKSC[+57]GERDVK	0.2*	1.8
G	T1000|S1004 TRAKSC[+57]GERDVKGIR	2.0**	1.4
	S1004 AKSC[+57]GERDVK	0.5*	0.8*	0.6	12.8****
H	S1059 ILTEDQDKIVR	2.8	2.9*
I	S1240 SLNVQLGR	1.0	0.8	0.9	0.7**

¹Peak areas for the indicated peptides were measured by PRM. Peak area ratio values of 1 indicates there was no difference detected between treatments, values >1 indicate an increase, and values<1 indicate a decrease in phosphopeptide peak area after treatment. Significance is included as follows

^*p≤0.05,

^**p≤0.01,

^***p≤0.001. and

^****p≤0.0001, Student's T-test.

²Phosphorylated residue is highlighted in bold.