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. 1999 Jan 1;13(1):35–48. doi: 10.1101/gad.13.1.35

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Copyright © 1999, Cold Spring Harbor Laboratory Press

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Expression of MMPs at the RNA and protein levels. (A) RT–PCR analysis of MMP transcripts in mRNA derived from TA3c cells. (Lane 1) Ladder of 100-bp DNA, ranging from 200–1500 bp; the top band corresponds to 2 kb; (lane 2) negative control PCR; (lane 3) MMP-2; (lane 4) MMP-3; (lane 5) MMP-7; (lane 6) MMP-9. (B) Zymograms of TA3c and TA3sCD44v6-10, TA3CD44tr, and TA3CD44H transfectant-derived supernatants, lysates, and crude membrane preparations. Gelatin (a,c,d) and β-casein (b) gels were used to test, respectively gelatinase A/MMP-2 (open arrow) and B/MMP-9 (solid arrow) and stromelysin (MMP-3, small arrows) activity in serum-free supernatants (a,b), water-soluble whole cell lysates (c) and 1% Triton-soluble crude membrane extracts of TA3 transfectants (d). Two independent isolates of each transfectant were tested. TA3 cells were transfected with the following: (Lanes 1,2) CD44tr; (lanes 3,4) CD44H; (lanes 5,6) sCD44v6-10; (lanes 7,8) sICAM-1; (lanes 9,10) sCD44v6-10R43A; (lanes 11,12), vector only. Molecular markers corresponding to 83 and 49 kD are indicated by arrowheads at left in a, c, and d; a molecular marker corresponding to 49 kD is indicated by an arrowhead at left in b. Bands corresponding to specific MMPs are shown.

Expression of MMPs at the RNA and protein levels. (A) RT–PCR analysis of MMP transcripts in mRNA derived from TA3c cells. (Lane 1) Ladder of 100-bp DNA, ranging from 200–1500 bp; the top band corresponds to 2 kb; (lane 2) negative control PCR; (lane 3) MMP-2; (lane 4) MMP-3; (lane 5) MMP-7; (lane 6) MMP-9. (B) Zymograms of TA3c and TA3sCD44v6-10, TA3CD44tr, and TA3CD44H transfectant-derived supernatants, lysates, and crude membrane preparations. Gelatin (a,c,d) and β-casein (b) gels were used to test, respectively gelatinase A/MMP-2 (open arrow) and B/MMP-9 (solid arrow) and stromelysin (MMP-3, small arrows) activity in serum-free supernatants (a,b), water-soluble whole cell lysates (c) and 1% Triton-soluble crude membrane extracts of TA3 transfectants (d). Two independent isolates of each transfectant were tested. TA3 cells were transfected with the following: (Lanes 1,2) CD44tr; (lanes 3,4) CD44H; (lanes 5,6) sCD44v6-10; (lanes 7,8) sICAM-1; (lanes 9,10) sCD44v6-10R43A; (lanes 11,12), vector only. Molecular markers corresponding to 83 and 49 kD are indicated by arrowheads at left in a, c, and d; a molecular marker corresponding to 49 kD is indicated by an arrowhead at left in b. Bands corresponding to specific MMPs are shown.