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Fig. 2 | BMC Cancer

Fig. 2

From: Inhibition of MMP-2 and MMP-9 decreases cellular migration, and angiogenesis in in vitro models of retinoblastoma

Fig. 2

Pharmacological inhibitors of MMP-2 and MMP-9 downregulate MMP2 and MMP9 mRNA. a The following MMPs were examined at the transcriptional level: MMP2, MMP7, MMP9, and MMP14. Y79 (left) and Weri-1 (right) cells were harvested for RNA isolation and cDNA synthesis. Material was pre-amplified using the TaqMan® PreAmp Master Mix with the respective primers. qPCR was done and results show mRNA expression relative to HPRT1 as endogenous control. Bar graphs indicate results ±SD; n = 3 biological replicates in triplicates. Y79 Rb cells express MMP2, MMP9 and MMP14; Weri-1 expressed MMP2 and MMP9. b-c Y79 (b) and Weri-1 (c) cells were treated with MMP-2 and MMP-9 inhibitors overnight. RNA and cDNA was extracted as in a showing that the inhibitors act at the transcriptional level. Bar graphs indicate fold change ±SD; n = 3. Standard deviation obtained from the biological replicates. d Knockdown of MMP2 and MMP9 by RNA interference shows on-target effects. Downregulation of MMP2 and MMP9 after siRNA compared to scramble samples. qPCR done as in a. e-f Reduction of MMP-2 and MMP-9 protein in Rb cells treated with MMPI (e) and siRNA (f); *p < 0.05, **p < 0.005. Western blot bar graphs indicate results ±SEM ratio of target protein to β-actin; n = 3. g-h ELISA analyses of MMP-2 and MMP-9 protein of whole cell lysates after treatment with MMPI (g) or siRNA (h); *p < 0.05, **p < 0.005. i-j, E2F regulates MMP expression in Y79 cells. Y79 cells treated with MMPI (i) or with siRNA (j) were assessed by Wb analysis for E2F. Western blot bar graphs indicate results ±SEM ratio of target protein to β-actin; n = 3; **p < 0.005

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