Skip to main content
. Author manuscript; available in PMC: 2009 Oct 27.
Published in final edited form as: J Invest Dermatol. 2008 May 8;128(10):2403–2411. doi: 10.1038/jid.2008.119

Figure 2. Gal3−/− keratinocytes are more sensitive to UVB-induced apoptosis in itro.

Figure 2

(a) Flow cytometric analysis of annexin-V stained keratinocytes. Keratinocytes from gal3+/+ and gal3−/− mice were untreated or irradiated with UVB at the indicated doses in vitro. Cells were harvested 24 h later and apoptosis was measured by flow cytometry with annexin-V-FITC and PI counterstaining. (b) Analysis of keratinocytes exposed to UVB for pycnotic nuclei. Keratinocytes were plated on coverslips and mock treated or irradiated with UVB at 100 J/m2, followed by incubation for 24 h. The cells were fixed, stained with Hoechst 33342, and analyzed for nuclear morphology of apoptosis by fluorescence microscopy. Cells with condensed or fragmented nuclei were determined as apoptotic cells. Data are expressed as percentage of apoptotic cells based on counting at least 150 cells. (c) Quantitative detection of histone-associated DNA fragments. Keratinocytes were irradiated with UVB (100 J/m2) or cultured with hydrogen peroxide (25 µM) or etoposide (25 µM). Twenty-four h later, histone-DNA complex was detected in an ELISA-based assay. Bars represent mean ± SD. *P<0.05. **P<0.001. Similar results were obtained in two additional experiments.