Journal: PLoS ONE

Article Title: TBCK Influences Cell Proliferation, Cell Size and mTOR Signaling Pathway

doi: 10.1371/journal.pone.0071349

Figure Lengend Snippet: HEK293 cells infected with the lentiviruses targeting to the various regions of TBCK mRNA (TBCK-RNAi-1 or 2) were subjected to Western blotting ( A ), cell counts ( B ), fluorescence microscopy ( C ) and MTT analyses ( D ). Bar, 200 µm. Lentivirus-infected cells were GFP-positive. Data are shown as the mean of three independent experiments ± SE (* P <0.05, ** P <0.01).

Article Snippet: Lentivirus-based RNA interference plasmid construction, lentivirus harvesting, and lentivirus titer testing were made by GENECHEM Company (Shanghai, China).

Techniques: Infection, Western Blot, Fluorescence, Microscopy

Journal: Aging (Albany NY)

Article Title: EFEMP2 indicates assembly of M0 macrophage and more malignant phenotypes of glioma

doi: 10.18632/aging.103147

Figure Lengend Snippet: WHO grade, IDH1 mutation and transcriptomic subtype preferences of EFEMP2 expression. ( A ) The correlation of EFEMP2 expression level with WHO grade. EFEMP2 expression levels in glioma of WHO grade II-IV in CGGA RNA-seq, TCGA RNA-seq, GSE16011 and REMBRANDT databases. *** p < 0.0001. ( B ) EFEMP2 expression in glioma specimens determined by IHC analysis. Scale bar, 60 μm. ( C ) The relationship between EFEMP2 transcription level and IDH1 mutation in CGGA and TCGA mRNA array datasets. *** p < 0.0001. ( D ) The relationship between EFEMP2 transcription level and transcriptomic subtype classification in CGGA and TCGA mRNA array datasets. ( E ) Correlation of EFEMP2 expression and IDH1 mutation in each transcriptomic subtype in CGGA and TCGA mRNA sequencing data, and GSE16011 dataset.

Article Snippet: The lentivirus constructing of overexpressing or knockdown EFEMP2 was obtained from GeneChem Co. Ltd. (Shanghai, PR, China).

Techniques: Mutagenesis, Expressing, RNA Sequencing, Sequencing

Journal: Aging (Albany NY)

Article Title: EFEMP2 indicates assembly of M0 macrophage and more malignant phenotypes of glioma

doi: 10.18632/aging.103147

Figure Lengend Snippet: EFEMP2 promotes GBM cell proliferation. ( A ) The expression of EFEMP2 was detected in three GBM cell lines by RT-qPCR. GAPDH was used as an internal reference. ( B ) RT-qPCR analysis of EFEMP2 expression in U251 cells overexpressing EFEMP2. Statistical significance was assessed using two-tailed Student’s t test. ***P < 0.001 (left). Western blotting (WB) analysis of EFEMP2 protein level in total cell lysates (T.C.L.) or conditioned media (C.M.) of cells with either vector or EFEMP2 stably overexpressed (right). ( C ) RT-qPCR analysis of EFEMP2 expression in U87 cells knocking down EFEMP2. Statistical significance was assessed using two-tailed Student’s t test. ***P < 0.001 (left). WB analysis of EFEMP2 protein level in total cell lysates (T.C.L.) or conditioned media (C.M.) of cells with either vector or EFEMP2 stably low expressed (right). ( D and E ) Proliferation of stable overexpressing ( D ) or knockdown ( E ) EFEMP2 cells as measured by EdU (green) uptake. Quantification of proliferation was measured by % EdU expressing cells / total cell number. Statistical significance was assessed using two-tailed Student’s t test. **P < 0.01. Scale bar, 60 μm. ( F and G ) The growth of cells with stable overexpressing ( F ) or knockdown ( G ) EFEMP2 was measured by Electric Cell-substrate Impedance Sensing (ECIS).

Article Snippet: The lentivirus constructing of overexpressing or knockdown EFEMP2 was obtained from GeneChem Co. Ltd. (Shanghai, PR, China).

Techniques: Expressing, Quantitative RT-PCR, Two Tailed Test, Western Blot, Plasmid Preparation, Stable Transfection, Knockdown, Electric Cell-substrate Impedance Sensing

Journal: Aging (Albany NY)

Article Title: EFEMP2 indicates assembly of M0 macrophage and more malignant phenotypes of glioma

doi: 10.18632/aging.103147

Figure Lengend Snippet: Patients with higher EFEMP2 transcription level exhibits poorer OS and PFS. ( A ) The half of patients with higher EFEMP2 expression exhibited shorter OS and PFS in Kaplan-Meier analyses based on CGGA mRNA array dataset. ( B ) The half of patients with higher EFEMP2 expression exhibited shorter OS and PFS in Kaplan-Meier analyses based on CGGA mRNA sequencing dataset. ( C ) Kaplan-Meier analyses of OS based on TCGA mRNA array, TCGA mRNA sequencing data, GSE16011 and REMBRANDT datasets. ( D ) The ROC curves indicating the sensitivity and specifcity of predicting 3- or 5-years of overall survival with EFEMP2 -level in CGGA, TCGA, or GSE16011 database.

Article Snippet: The lentivirus constructing of overexpressing or knockdown EFEMP2 was obtained from GeneChem Co. Ltd. (Shanghai, PR, China).

Techniques: Expressing, Sequencing

Journal: Aging (Albany NY)

Article Title: EFEMP2 indicates assembly of M0 macrophage and more malignant phenotypes of glioma

doi: 10.18632/aging.103147

Figure Lengend Snippet: Univariate and Multivariate Cox regression analyses and correlations with classic genetic alterations of EFEMP2. ( A – C ) Univariate and Multivariate Cox regression analyses of EFEMP2 expression level and several other clinical variables in CGGA and TCGA mRNA sequencing data, and GSE16011 data. ( D ) Correlations of EFEMP2 with classic genetic alterations of glioma. Grey background indicates wild-type or intact genes or chromosomes.

Article Snippet: The lentivirus constructing of overexpressing or knockdown EFEMP2 was obtained from GeneChem Co. Ltd. (Shanghai, PR, China).

Techniques: Expressing, Sequencing

Journal: Aging (Albany NY)

Article Title: EFEMP2 indicates assembly of M0 macrophage and more malignant phenotypes of glioma

doi: 10.18632/aging.103147

Figure Lengend Snippet: EFEMP2 is closely correlated with immune microenvironment of glioma. ( A ) Top 20 KEGG pathways derived from Gene Ontology analyses for EFEMP2 in CGGA and TCGA dataset. Thirteen KEGG pathways are overlap between the top 20 pathways of each dataset. ( B ) Pearson correlation analysis of EFEMP2 expression levels and immune responses in CGGA and TCGA datasets. Color depth and width of the bands represent the degrees of correlation. ( C ) Component types of the immune cells infiltrated into glioma are analyzed with CIBERSORT in CGGA and TCGA datasets.

Article Snippet: The lentivirus constructing of overexpressing or knockdown EFEMP2 was obtained from GeneChem Co. Ltd. (Shanghai, PR, China).

Techniques: Derivative Assay, Expressing

Journal: Aging (Albany NY)

Article Title: EFEMP2 indicates assembly of M0 macrophage and more malignant phenotypes of glioma

doi: 10.18632/aging.103147

Figure Lengend Snippet: EFEMP2 indicates assembly of M0 macrophage. ( A ) Correlation analysis of EFEMP2 and representative molecular of M0 ( CYP27A1 ), M1 ( IL12A , TNF ) and M2 ( IL13 , CCL22 , and MRC1 ) phenotype. ( B ) K-Means clustering (cluster = 3 or 4) based on whole genome expression profiling of M0, M1 and M2 phenotype from the dataset of a variety of resting and activated human immune cells (GSE22886) and CGGA RNA sequencing data. The samples named as “HIGH” were the top 20 samples with highest expression of EFEMP2 in CGGA. The samples named as “LOW” were the top 20 samples with lowest expression of EFEMP2 in CGGA. ( C ) K-Means clustering (cluster = 3 or 4) based on whole genome expression profiling of M0, M1 and M2 phenotype from the dataset of a variety of resting and activated human immune cells (GSE22886) and TCGA RNA sequencing data. The samples named as “HIGH” were the top 20 samples with highest expression of EFEMP2 in TCGA. The samples named as “LOW” were the top 20 samples with lowest expression of EFEMP2 in TCGA.

Article Snippet: The lentivirus constructing of overexpressing or knockdown EFEMP2 was obtained from GeneChem Co. Ltd. (Shanghai, PR, China).

Techniques: Expressing, RNA Sequencing