Oxidative Medicine and Cellular Longevity / 2018 / Article / Fig 6

Research Article

Intracellular and Intercellular Signalling Mechanisms following DNA Damage Are Modulated By PINK1

Figure 6

PINK1 loss of function impairs transmission of intercellular, bystander signalling. (a, b) Experimental scheme for the study of the bystander effect. For the direct treatment, the cells were exposed to BLM for 1 hour and then washed and incubated with fresh medium. This medium was conditioned for 24 hours with bystander factors by the directly treated cells and was used for the study of the bystander effect. The bystander cells are grown in medium transferred from BLM-treated donor cells collected at 24 h posttreatment. The transfer of media was performed between WT and KO MEFs and SC and KD SH-SY5Y as follows. The medium was transferred either from WT/SC to WT/SC and WT/SC to PINK1 KO/KD or from PINK1-KO/KD to WT/SC cells. (c, e) In MEFs, the MN induction and the number of γH2AX/53BP1 foci increase with the BLM concentration only when the WT cells are grown 24 h in medium transferred from WT cells. (d, f) For SH-SY5Y, the MN yield and γH2AX/53BP1 foci accumulated in both bystander SC and PINK1-KD cells grown in medium transferred from SC NB cells. Each data point represents the mean ± SEM of at least three independent experiments. values from one-way ANOVA are indicated in the figure.

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