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. 2017 Jun 27;8(47):81880–81891. doi: 10.18632/oncotarget.18737

Figure 3. 3T3-A-EXO promoted 3LL tumor cell invasion through MMP3-mediated increase in MMP9 activation.

Figure 3

(A) 3LL cells were treated with 30 μg/ml 3T3-EXO or 3T3-A-EXO for 4 h, and then the protein levels of uPA, MMP2, MMP3, MMP9 and cathepsin B were detected by Western blot. (B) After the treatment with 30 μg/ml 3T3-EXO or 3T3-A-EXO for 4 h, 3LL cells were collected and pre-treated with 20 nM UK 356618 for 2 h. Then, the invasive ability of these cells was measured by an in vitro invasive assay (n = 5). (C) 3LL cells were treated with 30 μg/ml 3T3-EXO or 3T3-A-EXO for 4 h. Then, cells were collected and cultured in serum-free RPMI 1640 media for an additional 24 h. MMP2 and MMP9 activity in the supernatants was detected by gelatin zymography. (D) 3LL cells were treated with 30 μg/ml 3T3-A-EXO for 4 h, and then the cells were collected and cultured in serum-free RPMI 1640 media in the presence of 10 nM UK 356618 for an additional 24 h. MMP2 and MMP9 activity in the supernatants was detected by gelatin zymography. E, 3LL cells were treated with 30 μg/ml 3T3-EXO or 3T3-A-EXO with or without 10 nM MMP9 inhibitor for 4 h, and then the invasive ability of these cells was measured using an in vitro invasive assay (n = 5). (A, C, D) One representative of three independent experiments is presented. (B, E) Results are presented as the mean ± SEM of three independent experiments. (B, D, E) DMSO is solvent control. P-values were generated by Student’s t-test or one-way ANOVA followed by Tukey-Kramer multiple comparisons test; *p< 0.05; ***p< 0.001; NS, not significant. Control indicates 3LL cells treated with PBS.