Further study indicated that FOXO promoted the expression of AMPs directly or indirectly, and enhanced the phagocytosis of bacteria in the shrimp

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Further study indicated that FOXO promoted the expression of AMPs directly or indirectly, and enhanced the phagocytosis of bacteria in the shrimp. element binding (CREB)-binding protein; JNK: c-JUN N-terminal kinase; ERK: extracellular regulated protein kinase; SIRT1: Sirtuin1.(TIF) ppat.1009479.s002.tif (5.1M) GUID:?8542D875-405D-4F20-9046-821015915E30 S3 Fig: Detection of the efficiency and off-target effects of RNAi in shrimp. (A) Efficiency of in in challenge analyzed by fluorescent immunocytochemical assay. Scale bar = 20 m. (E) Statistical analysis of FOXO nuclear translocation. WCIF ImageJ software was used to analyze co-localization by detecting the fluorescence intensity ratio of anti-FOXO (green) and DAPI-stained nuclei (blue) in hemocytes.(TIF) ppat.1009479.s003.tif (1.5M) GUID:?A5B05EB9-2632-4A14-8F49-9ED5E788BB12 Aloperine S4 Fig: Nucleotide sequence of promoter. The FOXO binding sites of genomic sequence were marked. Aloperine with green. Transcriptional start site marked with red.(TIF) ppat.1009479.s004.tif (161K) GUID:?82D15784-5B2F-4947-AC9E-1DE3B397D528 S5 Fig: RAB5 and RAB7 were upregulated in shrimp challenged with and play important role in shrimp antibacterial defense. (A, B) The mRNA expression patterns of (A) and (B), as analyzed by qPCR. (C, D) Expression patterns of RAB5 and RAB7 proteins was analyzed using western blotting. The results of statistical analysis of three replicates are shown in the lower panels of (C) and (D). The relative expression Aloperine levels of RAB5 or RAB7 normalized to that of -actin were expressed as the mean SD, and the value of the control shrimp was set as 1. (E) Efficiency of and RNAi, as determined using western blotting and qPCR. (F) The survival rate of shrimp after knockdown of or following infection. The survival rate of each group was calculated and the survival curves are presented as KaplanCMeier plots.(TIF) ppat.1009479.s005.tif (1.6M) GUID:?B586D3B7-E518-43A4-8B86-2A41C59358CC S6 Fig: FOXO promotes the expression RAB5 and RAB7 in intestine. (A) The mRNA expression level of the Aloperine and in intestine of and found that it was expressed at relatively basal levels in normal shrimp, but was upregulated significantly in shrimp challenged by and genes against bacterial infection. Furthermore, FOXO was identified as being Aloperine involved in cellular immunity by promoting the phagocytosis of hemocytes through upregulating the expression of the phagocytotic receptor scavenger receptor C (and and genes against bacterial infection. Furthermore, FOXO was identified as being involved in cellular immunity by promoting the phagocytosis of hemocytes through upregulating the expression of the phagocytotic receptor scavenger receptor C (and [3]. The transcriptional activity of FOXO is regulated by several kinds of post-translational modifications, such as phosphorylation, acetylation, methylation, and ubiquitination [4]. These modifications affect FOXO nuclear translocation or exit from the nucleus, and its interaction with co-repressors and co-activators, which can promote or decrease FOXO activity and mediate its different biological functions [5,6]. After activation, FOXO participates in many physiological functions by regulating the transcription of a variety of target genes [7]. FOXOs are activated by various extracellular stimuli, including growth factors, WASF1 cytokines, and hormones [8]. Growth signals, such as insulin or insulin-like growth factor 1 (IGF-1), interact with receptors of the insulin pathway, and activate phosphatidylinositol 3-kinase (PI3K) signaling, leading to phosphorylation of FOXO by the serine/threonine kinase, protein kinase B (AKT) [9]. Phosphorylated FOXO translocates from the nucleus to the cytosol, or prevents its translocation from cytosol to the nucleus, where it becomes ubiquitinated, leading to its degradation by the proteasome [10]. By contrast, in the absence of external growth signals, the PI3K-AKT signaling pathway is inactive, and unphosphorylated FOXO translocates into the nucleus to promote target gene transcription [11]. FOXO also participates in the anti-bacterial and anti-virus innate immune responses of invertebrates [12,13]. Chronic activation of FOXO in the aging intestines of represses the expression of peptidoglycan recognition protein SC2 (a negative regulator of the immune deficiency (IMD) pathway) and breaks.