E to cytotoxicity in THP-1 cells. It was also found responsible for higher levels of mature IL-1b. The NLRP3 inflammasome was found to mediate EHEC O157:H7-activated IL-1b production. Ehx may activate pro-caspase-1 through activation of NLRP3, like other pore-form bacteria toxins. However, the possibility that other types of inflammasome signaling may be activated by Ehx cannot yet be ruled out. This may also have stimulated the release of IL-1b. Cytotoxicity to THP-1 cells may also contribute to the release of IL-1b using some as yet unknown mechanism. Further study is needed to determine the possible roles of IL-1b in the pathogenesis of this potentially fatal foodborne infection.were infected with EDL933, DpO157, DehxA, or DehxA/pehxA. Cells were lysed over 2 h or 4 h postinfection mRNA expression of IL-1b was analyzed using RT-PCR. (TIF)AcknowledgmentsWe would like to thank Jennifer Cole of the Institute of Environmental and Human Health Texas Tech University for helping us to improve the English quality of this paper.Author ContributionsConceived and designed the experiments: JX XZ ZR YC. Performed the experiments: XZ YC YX HS HZ. Analyzed the data: XZ YC ZR CY HZ. Contributed reagents/materials/analysis tools: XZ YC YX CY HZ. Wrote the paper: XZ YC ZR JX.Supporting InformationFigure S1 mRNA expression of IL-1b in differentiatedTHP-1 cells. Differentiated THP-1 cells were left untreated or
Microtubules play an indispensable role in subcellular processes such as cell movement, 1379592 cell division and intracellular transportation. In turn, these processes are known to play a role in other biological phenomena such as wound healing, and cancer metastasis. Extracting information about the organization of microtubules in different cell lines could potentially shed light on the roles of microtubule associated proteins in that organization. While limited information is available about variation in microtubule distributions [1,2], information on those distributions in intact cells for different cell lines has not been readily available. Most microtubule BI-78D3 cost studies have focused on dynamics and interactions with drugs and microtubule associated proteins [3?6]. We believe that the ability to obtain reliable estimates of the overall organization of microtubules in whole cells could allow quantification of their dependency on different pertubagens, drugs, mechanical stimuli, etc.Electron microscopy can be used to trace microtubules, but the specimen preparation for imaging does not allow for intact cells to be imaged. Fluorescence microscopy can be used to image intact cells, but microtubules typically overlap and are often densely packed inside cells. It is very difficult, if not impossible, to Ornipressin manually trace each individual microtubule in a confocal or widefield fluorescence microscopy image in order to obtain accurate estimates of microtubule distribution parameters. Hence previous work comparing cell lines has often focused on the tips of microtubules where tracing is possible, or the comparison has been only qualitative [7]. We therefore previously developed an indirect method for estimating natural, interpretable and quantitative parameters such as the number and the mean length of microtubules from 3D fluorescence microscopy images of microtubules [8,9]. These parameters are important because they represent basic biophysical characteristics of tubulin polymerization. The basis of the method is to use a generative model of microtubule patterns (Fig.E to cytotoxicity in THP-1 cells. It was also found responsible for higher levels of mature IL-1b. The NLRP3 inflammasome was found to mediate EHEC O157:H7-activated IL-1b production. Ehx may activate pro-caspase-1 through activation of NLRP3, like other pore-form bacteria toxins. However, the possibility that other types of inflammasome signaling may be activated by Ehx cannot yet be ruled out. This may also have stimulated the release of IL-1b. Cytotoxicity to THP-1 cells may also contribute to the release of IL-1b using some as yet unknown mechanism. Further study is needed to determine the possible roles of IL-1b in the pathogenesis of this potentially fatal foodborne infection.were infected with EDL933, DpO157, DehxA, or DehxA/pehxA. Cells were lysed over 2 h or 4 h postinfection mRNA expression of IL-1b was analyzed using RT-PCR. (TIF)AcknowledgmentsWe would like to thank Jennifer Cole of the Institute of Environmental and Human Health Texas Tech University for helping us to improve the English quality of this paper.Author ContributionsConceived and designed the experiments: JX XZ ZR YC. Performed the experiments: XZ YC YX HS HZ. Analyzed the data: XZ YC ZR CY HZ. Contributed reagents/materials/analysis tools: XZ YC YX CY HZ. Wrote the paper: XZ YC ZR JX.Supporting InformationFigure S1 mRNA expression of IL-1b in differentiatedTHP-1 cells. Differentiated THP-1 cells were left untreated or
Microtubules play an indispensable role in subcellular processes such as cell movement, 1379592 cell division and intracellular transportation. In turn, these processes are known to play a role in other biological phenomena such as wound healing, and cancer metastasis. Extracting information about the organization of microtubules in different cell lines could potentially shed light on the roles of microtubule associated proteins in that organization. While limited information is available about variation in microtubule distributions [1,2], information on those distributions in intact cells for different cell lines has not been readily available. Most microtubule studies have focused on dynamics and interactions with drugs and microtubule associated proteins [3?6]. We believe that the ability to obtain reliable estimates of the overall organization of microtubules in whole cells could allow quantification of their dependency on different pertubagens, drugs, mechanical stimuli, etc.Electron microscopy can be used to trace microtubules, but the specimen preparation for imaging does not allow for intact cells to be imaged. Fluorescence microscopy can be used to image intact cells, but microtubules typically overlap and are often densely packed inside cells. It is very difficult, if not impossible, to manually trace each individual microtubule in a confocal or widefield fluorescence microscopy image in order to obtain accurate estimates of microtubule distribution parameters. Hence previous work comparing cell lines has often focused on the tips of microtubules where tracing is possible, or the comparison has been only qualitative [7]. We therefore previously developed an indirect method for estimating natural, interpretable and quantitative parameters such as the number and the mean length of microtubules from 3D fluorescence microscopy images of microtubules [8,9]. These parameters are important because they represent basic biophysical characteristics of tubulin polymerization. The basis of the method is to use a generative model of microtubule patterns (Fig.
