Effect on cell cycle and apoptosis after knocking down miR-21 in A-498 cells.

<p>(a) Effect on cell cycle: Flow cytometry analysis of the cell cycle showed a significant increase in G0/G1 phase (9.1%), with a concomitant reduction in the G2/M phase (6.8%), indicating that the forced reduction of miR-21 expression in A-498 cells could inhibit the cell growth and cause G0/G1 arrest (p value 0.010). (b) Effect on apoptosis: Apoptosis assay showed a marginal increase in the number of total apoptotic cells (5.17%) in the miR-21 anti-miRNA inhibitor transfected cells as compared to the negative control (1.79%) and mock (2.49%) (p value 0.017).</p

Correlation of miR-21 expression with 5 year survival of renal cell carcinoma patients.

<p>Correlation of miR-21 expression with 5 year survival of renal cell carcinoma patients.</p

<i>In vitro</i> and <i>In vivo</i> effect of genistein on the expression of miR-21.

<p>(a) Effect of genistein (25 µM) on the expression of miR-21 in A-498 cells: miR-21 expression was found to be reduced by ∼30% in genistein treated A-498 cells as compared to untreated cells. (b) <i>In vivo</i> effect of genistein on tumor size in nude mice: Genistein was able to reduce tumor formation significantly (black arrows, right panel).</p

Western analysis after the knock down of miR-21 in A-498 cells.

<p>Expression of p21, p38MAP kinase and cyclin E2 after the inhibition of miR-21 in A-498 cells, as compared to negative control. GAPDH was used as a normalizing control. p21 and p38 MAP kinase were up regulated, whereas cyclin E2 expression was found to be down regulated.</p

Effect on cell invasion and migration on A-498 cells after miR-21 inhibition.

<p>(a) Effect on cell invasion: Invasive properties of A-498 cells were decreased after the inhibition of miR-21 as compared to the negative control, Stained A-498 cells (Magnification-40×); Absorbance at 560 nm (p value 0.026). (b) Effect on cell migration: Migratory properties of A-498 cells were also decreased after the inhibition of miR-21 as compared to the negative control, Stained A-498 cells (Magnification-40×); Absorbance at 560 nm (p value 0.016).</p