Antitumor Macrophage Response to Bacillus pumilus Ribonuclease (Binase)

Figure 3

Quantification of phospho-p65 NF-κB protein (pp65) level in RAW 264.7 macrophages (a), (b) and THP-1-derived macrophages (c), (d) after 0.5 h (a), (c) or 3 h (b), (d) of treatment. Macrophages were treated for indicated time with RNAase1, binase, total RNA from A549 adenocarcinoma cells (10 μg/ml), and with RNA fragments obtained after RNA (500 μg/ml) hydrolysis by binase (2 ng/ml) or RNase1 (100 ng/ml) at 37°C, 300 rpm for 60 min. After, treatment cells were harvested and the protein level was measured using the Western blotting. Representative image of three independent experiments are shown. С: control (untreated cells); RNa1: RNase1 (10 μg/ml); Bi: binase (10 μg/ml); RfrR: RNA fragments, obtained after RNA hydrolysis by RNase1; RfrBi: RNA fragments, obtained after RNA hydrolysis by binase. Data represent mean ± SD (), , ns = nonsignificant.

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