Fig 2: IPMK?IECmice appear normal in homeostatic conditions. (A) IPMK was specifically depleted in intestinal epithelium using Villin-Cre mice. loxP sequences were inserted in the exon 6 region of the Ipmk gene, and Cre-driven deletion of flanked sequences resulted in truncated mRNA transcripts that were degraded readily. Tissue-specific IPMK knockout was confirmed at the (B) mRNA level and (C) protein level (n = 5, 3 per genotype in panel B). Red arrows indicate the IPMK band. Data are presented as means ± SD. (D) Body weights of 10-week-old mice showed no difference between genotypes (n = 6, 3 per genotype). Data are presented as means ± SD. (E) Gut permeability did not differ significantly between genotype in a normal state. Serum FITC-dextran concentration was measured 4 hours after a 400-mg/kg dose by oral gavage (n = 6, 6 per genotype). Data are presented as means ± SD. (F) No notable difference was observed in blots of mTOR complex and AKT signaling cascade. *P < .05, **P < .01. HSP90, heat shock protein 90; UTR, untranslated region.

Fig 3: Inhibition of LRP2 impairs insulin receptor internalization in C2C12 muscle cells.a Time course of IR internalization following insulin binding in C2C12 muscle cells. Cells were serum starved for 4 h and incubated with insulin as indicated, followed by sulfo-NHS-Biotin treatment. The biotinylated surface protein fraction was pull-downed (PD) with streptavidin-agarose beads. Immunoblot indicates total IR in biotin-labeled cell surface (top) and total lysates (bottom). Line graph shows the quantification of the relative cell surface IR amount at different time points by insulin treatment. These data are representative from two independent experiments. b IR internalization following insulin binding in C2C12 muscle cells lacking LRP2. Cells were transiently transfected by siRNA. Cells were starved for 4 h and incubated with insulin for 15 min, followed by sulfo-NHS-Biotin treatment. The biotinylated surface protein fraction was PD by streptavidin-agarose beads. Immunoblot indicates total IR in biotin-labeled cell surface (top) and total lysates (bottom). Bar graph shows the quantification of the relative cell surface IR amount in cell surface IR and internalized IR. These data are representative from two independent experiments. c Insulin-stimulated Akt phosphorylation in C2C12 muscle cells lacking LRP2. Cells were transiently transfected by siRNA. Cells were starved for 4 h and incubated with insulin for 15 min. Total cell lysates were separated by SDS–PAGE. Akt was visualized by immunoblotting. Graph shows the quantification of insulin-stimulated Akt phosphorylation from three independent experiments. d Insulin-stimulated glucose uptake in in C2C12 muscle cells lacking LRP2. Cells were transiently transfected by LRP2 siRNA. Cells were starved for 3 h and incubated with insulin for 30 min; n = 4 for control (no insulin), n = 6 control (insulin), n = 6 for LRP2 siRNA (no insulin), n = 5 for LRP2 siRNA (insulin). [3H]-2-deoxy-D-glucose uptake was measured. All bars and errors represent means ± SEM. p values by one-way ANOVA followed by post hoc Tukey’s HSD test are indicated. e Insulin-induced subcellular localization of FOXO1 in C2C12 cells. C2C12 muscle cells were treated with or without insulin in the presence of chlorpromazine or cytochalasin D. FOXO1 was detected by immunofluorescent analysis. These data are representative form three independent experiments. The scale bars represent 10 µm.

Fig 4: RMRP upregulates AKT expression by sponging miR-206. (A) Both RMRP and AKT are potentially targeted by miR-206 through the miRTarBase. (B) miR-206 mimics-induced inhibition of AKT can be completely restored by RMRP overexpression. (C) The luciferase reporter assay was performed to show that RMRP overexpression completely abrogates miR-206 mimics-induced inhibition of the reporter gene fused with the AKT 3'-UTR. **p < 0.01.

Fig 5: Flow cytometry of CD61 in mock and DENV infected MEG-01 cells. MEG-01 cells were mock-treated or treated with inhibitors of AKT (AKT-IV), PI3K (Ly294002), and PKCa (HBBDE). (A) Histograms from different conditions represent the expression of CD61 and (B) column graph presentation of median fluorescence intensity (MFI) of CD61 in cells with the above-stated condition. (*pValue < 0.05; ***pvalue < 0.001).