Figure 1
From: Fungus-derived hydroxyl radicals kill hepatic cells by enhancing nuclear transglutaminase
Cellular TG activity in HC cells increased, especially in the nucleus, upon co-incubation with pathogenic fungi. HC cells were seeded at 2 × 105 cells per well in a 6-well plate and incubated overnight. After adding 5-BAPA, the cells were incubated (a) alone (row 1) or were co-incubated with 2.5 × 106 C. albicans cells (row 2) or S. cerevisiae cells (row 3) for 24 hours; (c) with different doses of C. albicans cells for 24 hours; (d) with 5 × 106 C. albicans cells for the indicated time; (e) with 5 × 106 C. albicans cells in the absence (row 2) and presence of 100 µM of TG2 inhibitors, cystamine (row 3) or R283 (row 4) for 24 hours; (g) alone (row 1) or were co-incubated with either 5 × 106 C. albicans cells (row 2) or the same number of C. glabrata cells (row 3) in an insert cup with a 0.4-µm pore size; (i) alone (row 1) or were co-incubated with living 5 × 106 (row 2) or heat-killed 5 × 109 (row 3) C. albicans cells for 24 hours; or (k) alone (row 1) or were co-incubated with living 5 × 106 C. glabrata (row 2), C. utilis (row 3) and Schizo. pombe cells (row 4) for 24 hours. Scale bars = 20 µm. Representative images from at least 3 fields from 3 independent experiments are shown for (a,e,I and k) and from at least 3 fields from a single experiment for (g). Fluorescence intensities from TRITC in both the cytoplasm and nucleus of panels (b,c,d,f and j) were quantitated using ZEN 2011 software, and relative TG activity levels in both locations are presented in bar graphs, with the levels from HC cells incubated alone (panels b,c,f,h and j) or at time 0 (d) used as the controls (mean ± SD, n = 3). Fluorescence intensities from Alexa 488 signal from panel (h) were quantitated using ZEN 2011 software, and the mean ± SD are presented in bar graphs after subtraction of background intensities. Asterisks represent statistical significance.
