By visualizing DNA with diamidino phenylindole (DAPI), we found that hypothermal incubation followed by rewarming of human neutrophils resulted in an increased number of DAPI-positive objects representative of extensi...By visualizing DNA with diamidino phenylindole (DAPI), we found that hypothermal incubation followed by rewarming of human neutrophils resulted in an increased number of DAPI-positive objects representative of extensive DNA unfolding seemingly similar to neutrophil extracellular traps (NETs). In contrast to canonical NET formation, diphenylene iodonium (DPI), an NADPH oxidase inhibitor, exhibited negligible effects on formation of the DAPI-positive objects. Moreover, multiple instances of DNA damage were detected in the objects, but not in canonical NETs. Our results thus suggest the potential of hypothermia for triggering DNA structural alteration in neutrophils, which is similar to but distinct from NET formation.展开更多
While micronuclei (MN) store extranuclear DNA and cause genome instability, the effects of nuclear envelope (NE) assembly defects associated with MN on genome instability remain largely unknown. Here, we investigated ...While micronuclei (MN) store extranuclear DNA and cause genome instability, the effects of nuclear envelope (NE) assembly defects associated with MN on genome instability remain largely unknown. Here, we investigated the NE protein distribution in MN using HeLa human cervical cancer cells. Under the standard condition and two pharmacological culture conditions, we found that three types of NE protein assemblies were associated with MN: 1) intact NE assembly, in which both core and non-core NE proteins were evenly present;2) type I assembly, in which only core NE proteins were detectable;and 3) type II assembly in which a region deficient for both core and non-core NE proteins existed and a pattern recognition receptor, cyclic guanosine monophos-phate-adenosine monophosphate synthase, was frequently detected. Our findings provide experimental settings and a method of grouping MN-associated NE defects, which may be helpful for researchers who are interested in regulation of genome and nuclear organization relevant to cancer development.展开更多
文摘By visualizing DNA with diamidino phenylindole (DAPI), we found that hypothermal incubation followed by rewarming of human neutrophils resulted in an increased number of DAPI-positive objects representative of extensive DNA unfolding seemingly similar to neutrophil extracellular traps (NETs). In contrast to canonical NET formation, diphenylene iodonium (DPI), an NADPH oxidase inhibitor, exhibited negligible effects on formation of the DAPI-positive objects. Moreover, multiple instances of DNA damage were detected in the objects, but not in canonical NETs. Our results thus suggest the potential of hypothermia for triggering DNA structural alteration in neutrophils, which is similar to but distinct from NET formation.
文摘While micronuclei (MN) store extranuclear DNA and cause genome instability, the effects of nuclear envelope (NE) assembly defects associated with MN on genome instability remain largely unknown. Here, we investigated the NE protein distribution in MN using HeLa human cervical cancer cells. Under the standard condition and two pharmacological culture conditions, we found that three types of NE protein assemblies were associated with MN: 1) intact NE assembly, in which both core and non-core NE proteins were evenly present;2) type I assembly, in which only core NE proteins were detectable;and 3) type II assembly in which a region deficient for both core and non-core NE proteins existed and a pattern recognition receptor, cyclic guanosine monophos-phate-adenosine monophosphate synthase, was frequently detected. Our findings provide experimental settings and a method of grouping MN-associated NE defects, which may be helpful for researchers who are interested in regulation of genome and nuclear organization relevant to cancer development.
