1. Evidence for an exclusive association of matrix metalloproteinase-9 with dysfunctional high-density lipoprotein: A novel finding 
S. Sini, D. Deepa, S. Harikrishnan, N. Jayakumari 
Atherosclerosis 
Volume 236, Issue 1, Pages (September 2014)
DOI: /j.atherosclerosis
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2. Fig. 1
Serum gelatin zymography. Gelatin zymography was employed to assess the activity of serum MMP-2 and MMP-9.Serum (10 μl, 1:200 diluted) was loaded on 7.5% SDS-PAGE gel. Inset – representative gelatin zymogram gel and the gelatinolytic activity detected as transparent bands. Lane 1 & 2- serum from healthy subjects and lane 3, 4 & 5- serum from CAD patients. The graph represents the densitometric values. Values are the mean of 30 samples ±SD, ‘*’p < 0.05.
Atherosclerosis  , DOI: ( /j.atherosclerosis )
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3. Fig. 2
MMP 9 activity in isolated lipoproteins: HDL- both functional and dysfunctional and apo B containing lipoproteins were isolated by UC and assessed for gelatinase activity using zymography as described in 2.4. (a) Inset shows the representative zymogram. Lane 1 & 2- functional HDL, lane 3, 4, 5 & 6- dysfunctional HDL. Densitrometric values are represented graphically. Values are the mean ± SD for 10 separate experiments. ***p <  functional-vs. dysfunctional HDL (b): Representative zymmogram of serum lipoprotein fractions from subjects having dysfunctional HDL. Lane 1- apoB containing lipoproteins [VLDL, LDL and Lp(a)] .Lane 2- serum without apo B containing lipoproteins and with HDL. Lane 3- lipoprotein-free serum [after separation of apoB and HDL], lane 4- dysfunctional HDL fraction.
Atherosclerosis  , DOI: ( /j.atherosclerosis )
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4. Fig. 3
MMP 9 activity in HDL fractions isolated by three different methods and corresponding functionality of HDL. (a) Western blot of HDL fractions. Apo A1 was used as internal control. Lane 1, 2, 3 dysfunctional HDL fractions isolated by UC(1), PEG(2) and EE(3)methods. Lane 4- functional HDL [UC]. (b) dot blot assay of EE HDL. Serum was subjected to PAGE in 3.75% gel. The band corresponding to HDL was excised, electroeluted and probed with MMP9 antibody. Lane 1&2- functional-and dysfunctional HDL respectively. (c) Gelatin zymogram of HDL isolated by different methods. Lane 1- functional HDL [UC], lane 2, 3, 4 dysfunctional HDL isolated by (2)UC, (3)PEG, (4)EE methods. (d) Antioxidant property of HDL. HDL isolated by three different methods. Values are the mean of 10 experiments ±SD, ‘***’p < 
Atherosclerosis  , DOI: ( /j.atherosclerosis )
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5. Fig. 4
MMP 9 activity in HDL fraction pre-incubated with MMP9 and functionality of HDL. Functional HDL was treated with MMP9 (a) Western blot of HDL incubated with MMP-9. (a) lane 1- functional HDL without MMP-9 treatment, lane 2- functional HDL treated with MMP-9. (b) lane 1- functional HDL treated with MMP9 alone, lane 2- HDL after treatment with MMP9 exposed to EDTA (10 mM). (c) Gelatin zymogram of HDL treated with MMP-9. Lane 1-functional HDL without MMP9, lane 2- HDL treated with MMP-9. (d) Lane 1-HDL treated with MMP9 alone, lane 2- HDL after treatment with MMP9 exposed to EDTA (10 mM). (e) Functional property of HDL treated with MMP-9. LDL was subjected to air oxidation with and without native functional HDL or HDL treated with MMP9 values are the mean of 10 experiments ±SD, ‘***’p < 
Atherosclerosis  , DOI: ( /j.atherosclerosis )
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6. Fig. 5
Proteolytic activity of MMP 9 on Apo A1 .Functional HDL fractions were incubated with MMP 9 standard for 24 h as described in methodology. Aliquots were electrophoresised in 10% SDS-PAGE. Lane 1, 3- functional HDL fraction without MMP 9, lane 2, 4- same HDL fraction treated with MMP 9.
Atherosclerosis  , DOI: ( /j.atherosclerosis )
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7. Fig. 6
MMP 9 activity in HDL subfractions. HDL and its major subfractions-HDL2 (density1.063–1.125) and HDL3 (density 1.125–1.21) were isolated by ultracentrifugation and desalted. HDL fractions were subjected to western blot and gelatin zymography. (a) Western blot of HDL subfractions: lane 1- dysfunctional HDL, lane 2 &3- HDL2 and HDL3 subfractions of the same dysfunctional HDL. (b) Gelatin zymogram of HDL and its subfractions. Lane 1- dysfunctional HDL, lane 2 and 3- HDL3 and HDL2 subfractions of the same dysfunctional HDL showing gelatinolytc activity.
Atherosclerosis  , DOI: ( /j.atherosclerosis )
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8. Fig. 7
Monocyte/macrophage cytokine release induced by HDL. Cells (1 × 106 cells/ml) were cultured under standard condition and treated with functional HDL, dysfunctional HDL, MMP 9 standard, MMP9 treated HDL or MMP9 treated HDL after exposure to EDTA, for 24 h as described in methods. Levels of cytokines- TNF-α and IL-10, in the culture medium were quantitated using respective ELISA kits. Values are the mean of 10 independent experiments ±SD, ‘***’p < 
Atherosclerosis  , DOI: ( /j.atherosclerosis )
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