Supplementary MaterialsFigure S1: Manifestation of prohibitin 2 (PHB2) in PHB2-overexpressing embryonic stem (ES) cells and PHB2-knockdown ES cells. Similarly, the EBRTcH3 cells transporting the under the control of a Tc-regulated promoter were cultured with (S4) or without (S3) Tc for 4 days, and subjected to time laps analysis using a confocal microscope. gene and are expressed in the absence of Tc. Level pub, 5 m.(AVI) pone.0081552.s003.avi (3.6M) GUID:?D2602167-4533-479E-BCCA-20973465C787 Movie S3: Time EDC3 laps analysis of mitochondria in ES cells. EBRTcH3 cells transporting the gene under the control of a Tc-regulated promoter were cultured with Tc (S2) or without Tc (S1) for 4 days, and subjected to time laps analysis using a confocal microscope. Similarly, the EBRTcH3 cells transporting the under the control of a Tc-regulated promoter were cultured with (S4) or without (S3) Tc for 4 days, and subjected to time laps analysis using a confocal microscope. gene and are expressed in the absence of Tc. Level pub, 5 m.(AVI) pone.0081552.s004.avi (3.1M) GUID:?96F61AA2-E01B-4EF7-834F-65B106EE7CB3 Movie S4: Time laps analysis of mitochondria in ES cells. EBRTcH3 cells transporting the gene under the control of a Tc-regulated promoter were cultured with Tc (S2) or without Tc (S1) for 4 days, and subjected to time laps analysis using a confocal microscope. Similarly, the EBRTcH3 cells transporting the under the control of a Tc-regulated promoter were cultured with (S4) or without (S3) Tc for 4 days, and subjected to time laps analysis using a confocal microscope. gene and are expressed in the absence of Tc. Level pub, 5 m.(AVI) pone.0081552.s005.avi (4.0M) GUID:?5AA5934E-70B4-4C64-870D-9F4073AA3F5E Abstract Background The pluripotent state of embryonic stem (ES) cells is definitely controlled by a network of specific transcription factors. Recent studies also suggested the significant contribution of mitochondria within the rules of pluripotent stem cells. However, the molecules involved in these regulations are still unfamiliar. Strategy/Principal Findings With this study, we found that prohibitin 2 (PHB2), a pleiotrophic element primarily localized in mitochondria, is definitely a crucial regulatory element for the homeostasis and differentiation of ES cells. PHB2 was highly indicated in undifferentiated mouse ES cells, and the manifestation was decreased during the differentiation of ES cells. Knockdown of PHB2 induced significant apoptosis in pluripotent ES cells, whereas enhanced manifestation of PHB2 contributed to the Resminostat hydrochloride proliferation of ES cells. However, enhanced manifestation of PHB2 strongly inhibited ES cell differentiation into neuronal and endodermal cells. Interestingly, only PHB2 with intact mitochondrial focusing on signal showed these specific effects on ES cells. Moreover, overexpression of PHB2 enhanced the processing of a dynamin-like GTPase (OPA1) that regulates mitochondrial fusion and cristae redesigning, which could induce partial dysfunction of mitochondria. Conclusions/Significance Our results suggest that PHB2 is Resminostat hydrochloride definitely a crucial mitochondrial regulator for homeostasis and lineage-specific differentiation of ES cells. Intro The pluripotent stem cells, such as embryonic stem (ES) Resminostat hydrochloride cells and induced pluripotent stem (iPS) cells, are controlled by a specific transcription network composed of core transcription factors such as Oct4, Resminostat hydrochloride Sox2, and Nanog [1], [2]. Recent reports suggested the involvement of other factors, such as mitochondrial functions, in the rules of stem cells [3], [4]. Previously, we performed proteomic analyses of mouse ES cells and recognized prohibitin 2 (PHB2) as one of the highly indicated proteins in pluripotent mouse ES cells [5]. PHB2 is a pleiotropic protein that has been reported to be essential for cell proliferation and development in higher eukaryotes [6], [7], [8]. PHB2 is mainly involved in the functionality of the mitochondrial inner membrane like a protein-lipid scaffold. Some reports also suggested additional functions of PHB2, such as transcriptional rules in the nucleus and cell signaling in the plasma membrane [9]. Recent studies suggested numerous tasks of PHBs in disease pathogenesis. For example, PHBs are involved.