5 pretreatment and B P seems to be the most efficient. Note that the particle coupled PAH are bioavailable in our system since CYP1A1 mRNA and its enzymatic activity together were increased. Moreover, when different light PAH found on particles were tested, the antiapoptotic effect was not found. We also took into consideration the effect of biological compounds adsorbed onto particles, such as endoto ines, by using a specific bacteria LPS neutraliz ing protein rENP. This did not diminish the protector effect of PM2. 5 from apoptosis induced by A23187 and STS indicating that endo to ins are not involved in the process. Altogether, our data strongly suggest that water soluble and heavy PAH components contribute to the antiapoptotic effect of Parisian PM2. 5 observed in human bronchial epithelial cells.

The antiapoptotic mechanism is mediated by the aryl hydrocarbon receptor To delineate the molecular mechanism of the antiapop totic effect of PM2. 5 efficient at the mitochondrial checkpoint, we focused on the aryl hydrocarbon recep tor activated after cell e posure to organic com pounds such as PAH. Indeed, AhR is a ligand induced transcription factor which relocates to the nucleus and induces the e pression of numerous target genes. Thus, we investigated the possible implication of AhR in our process. To test this we first either activated or inhibited AhR, using an agonist or an antagonist. Figure 7A shows that beta NF used prior to A23178 significantly reduced the amount of apoptotic cells low and further improved the protection conferred by PM2. 5 e posure low.

Conversely, pretreatment with alpha NF significantly reduced the protection pro vided by PM2. 5 e posure low although it did not noticeably modify the apoptotic effect of A23187. These findings are con sistent with the involvement of AhR in the antiapoptotic effect of PM2. 5 e posure. Finally, we tested the effect of AhR silencing in the antiapoptotic effect observed after PM2. 5 e posure. For this, we used validated fluorescent siRNA in order to select the fluorescent positive cells by flow cytometry. After siRNA optimization and validation of AhR silencing by western blot, DiOC 3 and PI assays were performed by flow cytometry on cells e posed or not to PM2. 5 and or A23187 for 24 h as before. Figure 7B shows that AhR silencing significantly reduced the protection triggered by PM2.

5 3 low alike the antagonist did. Interestingly, both the AhR silencing and AhR antagonist partially reduced the PM2. 5 protective effect with almost the same e tent. The increase in alpha NF concentration or siRNA AhR amount did not completely abolish the protection suggesting that another pathway might be involved. Taken together, these results suggest that AhR Dacomitinib partially contributes to the antiapoptotic effect of PM2. 5 e posure.