Fig. 2

Dox-induced DSBs elicit a rapid NPM secretion in the absence of necrosis. a Representative WB using γH2AX and NPM antibodies showed that γH2AX levels were induced upon 1μM Dox in hCmPCs after 4 and 8h of treatment while NPM levels were not modulated. Beta-actin was used as loading control. b Densitometric analysis of γH2AX expression levels normalised by β-actin (n = 3;**P < 0.01;***P < 0.001). c Densitometric analysis of NPM expression levels normalised by β-actin (n = 3). Uncropped images of blots are shown in Additional file 1: Figure S1b. d hCmPCs were treated for 4 h and 8 h with 1 μM Dox, serum-free media were collected and analysed by ELISA assay for NPM expression. NPM was induced at 8h with respect to 4h treatment both in Ctrl- and Dox-treated cells. The difference at 8h between Dox and Ctrl was statistically significant (n = 7;*P < 0.05). e LDH assay, a marker of cell necrosis, showed that the release does not depend on a passive release from dead cells (n = 6). A negative control and positive control were inserted in the assay; the latter is statistically significant vs negative control (n = 6; ^*P < 0.05)