Fig. 1

siRNA suppression of TIMP-2 in the OVCAR5 cell line. Suppression of TIMP-2 expression by siRNA transfection in the OVCAR5 cell line is described in “Methods”. TIMP-2 expression was evaluated by immunofluorescence at the protein level and at mRNA level by qRT-PCR as described in “Methods”. A, B and C are single siRNA duplexes and A + B + C is representative of a pool of all three TIMP-2 siRNAs at a 3 nM final concentration. Immunofluorescence images are representation of merged DAPI (blue) and TIMP-2 (red) staining on individual cell lines done in three passages in triplicate. The intensity of fluorescence was obtained using FIJI software. ×20 magnification; scale bar (in yellow) 20 μM; P indicates the parental cell line treated with transfection reagent only, Cont are cells transfected with scrambled siRNA. For mRNA expression, graphs represent amount of mRNA relative to 18S + SEM derived from three experiments done in triplicate. Significance was determined by one-way ANOVA and indicated by ***p < 0.001, ****p < 0.0001