Fig 1: Downregulation of AD-associated proteins in mice lacking the oligodendrocyte network key driver Cnp. a Representative western blots of BIN1, GOT2, CNP and alpha-TUBULIN stainings on Cnp-WT and Cnp-KO corpus callosum (CC) tissues (n = 3). b Quantification of BIN1 and GOT2 protein expression in Cnp-KO corpus callosum compared to Cnp-WT corpus callosum (protein expression normalized to alpha-TUBULIN expression for each sample, n = 3 per condition; p = 0.0041, p = 0.0007, p = 0.032). c Representative confocal image of post-natal day 60 sections of Cnp-WT and Cnp-KO corpus callosum, stained for OLIG2 (red), BIN1 (green) and DAPI (blue), scale bar = 300 µm. The bottom panel shows higher magnification of the double stainings (white arrowheads indicate double positive for BIN1 and OLIG2), scale bar = 50 µm. d Quantification of the proportion of each cell types expressing BIN1 in Cnp-WT and Cnp-KO corpus callosum (oligodendrocytes – OLIG2+ in gray, neurons – NeuN+ in white, astrocytes – GFAP+ in black; n = 3–4). e Quantification of the percentage of OLIG2+ cells expressing BIN1 in Cnp-WT and Cnp-KO corpus callosum (n = 3–4). f Representative confocal image of post-natal day 60 sections of Cnp-WT and Cnp-KO cortex, stained for OLIG2 or NeuN (red), GOT2 (green) and DAPI (blue), scale bar = 300 µm. The bottom panel shows magnification of the double stainings (white arrowheads indicate double positive for GOT2 and OLIG2 or NeuN, respectively), scale bar = 50 µm. g Quantification of the proportion of each cell types expressing GOT2 in Cnp-WT and Cnp-KO cortex (oligodendrocytes – OLIG2+ in gray, neurons – NeuN+ in white, astrocytes – GFAP+ in black; n = 3–4). h Quantification of the percentage of OLIG2+ cells and of NeuN+ cells expressing GOT2 in Cnp-WT and Cnp-KO corpus callosum (n = 3–4). * p < 0.05, ** p < 0.01, *** p < 0.001
Fig 2: Flow Chart. #The associations of the genes with liver cancer prognosis that were not previously reported. *By text mining of searching for key words related to the markers of liver cancer prognosis and screening, a total of 1,173 genes containing both previously reported and unreported associations with liver cancer prognosis were obtained. **In each sample, the simultaneous high expression of all three genes was considered as high expression group in new combination. Similarly, the simultaneous low expression of all three genes was considered as low expression group in new combination. ***By comparing the prognostic value of individual genes and their combinations, we selected genes of combination F2- GOT2-TRPV1 for subsequent verification.
Fig 3: The expression of F2, GOT2, and TRPV1 in 90 pairs of HCC and adjacent normal liver tissues of biological tissue microarray by IHC, and the association with HCC patients prognosis. (A) Negative, weakly positive, intermediately positive, and strongly positive IHC staining of F2, GOT2, and TRPV1. F2, GOT2, and TRPV1 were all low expressed in liver cancer. (B) The lower protein expression levels of F2, GOT2, and TRPV1 were all associated with 5-year OS of 90 HCC patients, examing by Kaplan-Meier analyses and log-rank test. However, there was marginally significant association between the F2-GOT2-TRPV1 combination protein expression levels with the OS of HCC patients. (F2: p = 0.033, GOT2: p = 0.035, TRPV1: p = 0.046, F2-GOT2-TRPV1: p = 0.051).
Fig 4: Expression of F2, GOT2, and TRPV1 in HCC and adjacent normal liver tissues confirmed by independent microarrays from the Oncomine database. The expression of (A) F2, (B) GOT2, and (C) TRPV1 were all significantly reduced in HCC tissues by the Roessler Liver 2 Statistics [225 HCC tissues (dark blue) vs. 220 normal liver tissues (light blue)]. ***p < 0.001.
Fig 5: The relative expression levels of F2, GOT2, and TRPV1 were assessed by qRT-PCR in HCC tissues and adjacent liver tissues. (A) The expression levels of F2 were assessed by qRT-PCR in 20 pairs of HCC tissues and peritumoral tissues. Paired t-test, ***p < 0.001. (B) The relative expression levels of GOT2 were assessed by qRT-PCR in 19 pairs of HCC tissues and peritumoral tissues. Paired t-test, ***p < 0.001. (C) The relative expression levels of TRPV1 were assessed by qRT-PCR in 19 pairs of HCC tissues and peritumoral tissues. Paired t-test, log, **p < 0.01.
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