Generation of Adducts of 4-Hydroxy-2-nonenal with Heat Shock 60 kDa Protein 1 in Human Promyelocytic HL-60 and Monocytic THP-1 Cell Lines

Figure 1

Confocal and epifluorescence microscopic analysis of HL-60 cells treated with 10 μM HNE for 2 h. HL-60 cells (5 × 10⁴) were harvested after 2 h of treatment with 10 μM HNE, fixed, permeabilized, blocked, and incubated overnight with the anti-HNE-histidine primary antibody at 4°C, as described in Section 2. Cells were then washed and exposed to the FITC-conjugated secondary antibody for 1 h at rt, washed again, mounted on coverslips, and examined under a Leica TCS SP2 confocal microscope, equipped with a water immersion HCX Apo 0.8 objective (a). For epifluorescence microscope analysis (Axioskop, Carl Zeiss), slides were also counterstained with DAPI (b).

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(b)