*2.4. Immunofluorescence*

Tissue sections were deparaffinized and rehydrated. Heat-induced antigen retrieval in 10mM sodium citrate, pH 6.0, 0.05% Tween 20 was done prior to immunostaining for 4-hydroxynonenal (4-HNE) protein adducts and non-receptor tyrosine kinase SRC (SRC), but not for 8-hydroxyguanosine (8-OHG). The slides were washed in Tris Buffered Saline 1×, 0.02% Tween 20 (TBS-T) and were incubated in TBS-T, 5% normal goa<sup>t</sup> serum (NGS) at room temperature for 30 min. Tissue sections were incubated overnight at 4 ◦C with the primary antibodies diluted in TBS-T, 1% NGS (Table 1). They were washed twice in TBS-T and incubated at room temperature for 1 hour with the secondary antibodies diluted in TBS-T, 1% NGS, 1 μg/mL4',6-diamidine-2'-phenylindole dihydrochloride (DAPI). After one wash in TBS-T and two washes in TBS 1X, slides were mounted with PermaFluor™ mounting medium (ThermoFisher Scientific, Montreal, QC, Canada). For the negative controls, the primary antibodies were omitted.


**Table 1.** List of antibodies used for immunofluorescence experiments.

8-hydroxyguanosine (8-OHG), 4-hydroxynonenal (4-HNE), proto-oncogene tyrosine-protein kinase (SRC).
