Journal: PLoS ONE

Article Title: Candidate Genes for Expansion and Transformation of Hematopoietic Stem Cells by NUP98-HOX Fusion Genes

doi: 10.1371/journal.pone.0000768

Figure Lengend Snippet: Genes changed by NA10 and ND13 but not by the ND13(N51S) mutant compared to the GFP control.

Article Snippet: The following genes were analyzed (TaqMan assay ID number is indicated, Applied Biosystems); HOXA5 (Hs00430330_m1), HOXA7 (Hs00600844_m1), HOXA9 (Hs00365956_m1), HOXA10 (Hs00538183_m1), PBX1 (Hs00231228_m1), PBX2 (Hs00855025_s1), PBX3 (Hs00608415_m1), MEIS1 (Hs00180020_m1), MEF2C (Hs00231149_m1), JAG2 (Hs00171432_m1), PRNP (Hs00175591_m1), DDX4 (Hs00251859_m1) and HLF (Hs00171406_m1).

Techniques: Mutagenesis, Control, Transduction, Binding Assay, Coagulation, Cell Differentiation, Membrane

Journal: Cancer Cell International

Article Title: The expression of pro-prion, a transmembrane isoform of the prion protein, leads to the constitutive activation of the canonical Wnt/β - catenin pathway to sustain the stem-like phenotype of human glioblastoma cells

doi: 10.1186/s12935-024-03581-1

Figure Lengend Snippet: Prognostic significance of PrP C expression in GBM and transcript levels in patient-derived GSC cultures. Kaplan–Meier survival analysis of overall survival and disease-free survival of low-grade gliomas (LGG) (total 514 patients) ( A ) and GBM (total 162 patients) ( B ) with high or low expression level of PRNP , using GEPIA ( http://gepia2.cancer-pku.cn/#index ) in TCGA database ( https://portal.gdc.cancer.gov/ ). A statistically significant difference was observed only in GBM (OS: P = 0.014; PFS: P = 0.026). C) PRNP transcript levels in GSCs isolated from 15 human GBMs. Data was obtained by RNA-seq, and expressed as counts per million reads mapped (cpm)

Article Snippet: GSCs were also transfected with human PRNP natural open reading frame (ORF) mammalian expression plasmid (Sino Biological Inc.), to stably overexpress PrP C (GBM OV); the same plasmid was used to revert PrP C down-regulation in GBM KD cells (GBM REV).

Techniques: Expressing, Derivative Assay, Isolation, RNA Sequencing Assay

Journal: Prion

Article Title: Differentiated cultures of an immortalized human neural progenitor cell line do not replicate prions despite PrP C overexpression

doi: 10.1080/19336896.2023.2206315

Figure Lengend Snippet: Summary of ReN cell lines. WT – wild type; KO – PRNP knocked out; NPC – neural progenitor cell; PuroR – puromycin resistance; GFP – green fluorescence protein.

Article Snippet: The following TaqMan gene expression assays were used: ACTB (Hs03023943_g1), GFAP (Hs00909233_m1), TUBB3 (Hs00801390_s1), PRNP (Hs01920617_s1) and PRNP_CDS (Hs04937277_s1).

Techniques: Fluorescence, Sequencing, Expressing

Journal: Prion

Article Title: Differentiated cultures of an immortalized human neural progenitor cell line do not replicate prions despite PrP C overexpression

doi: 10.1080/19336896.2023.2206315

Figure Lengend Snippet: Lentiviral-transduced ReN cell lines overexpress PrP C and differentiate into neurons and astrocytes. (a) Lentiviral-transduced ReN cell lines were produced that overexpress the mouse, hamster and human 129M and 129V versions of PrP C . Lysate from proliferating ReN progenitor cell lines was western blotted for PrP C using the 6H4 (recognizes mouse and human PrP) and 3F4 (recognizes hamster and human PrP) monoclonal antibodies. Total protein signal was measured in Bio-Rad TGX stain-free gels according to manufacturer’s instructions. (b) the 129M and WT ReN cell lines exhibit a similar pattern of protein expression for PrP, TUBB3 and GFAP throughout the process of differentiation into neurons and astrocytes. ReN cells were differentiated as thin-3D cultures and lysate collected at weekly timepoints post-differentiation was western blotted for PrP (via 3F4 mAb), TUBB3 and GFAP. Signal was normalized to the most prominent band in the corresponding total protein image (see arrow). (c) Normalized protein expression of PrP, TUBB3 and GFAP in ReN WT and 129M cells throughout standard differentiation. Signal intensity was quantified using imageJ and normalized to the signal from the strongest band identified in total protein images. (d) qPCR-based quantification of GFAP, TUBB3 and PRNP of ReN 129M, WT and Empty cells over 4 weeks of differentiation. PRNP was quantified using PCR primers that bind the non-coding mRNA region (PRNP) and protein-coding region of the mRNA (PRNP_CDS). * p -value<0.05 (calculated using one-way ANOVAs).

Article Snippet: The following TaqMan gene expression assays were used: ACTB (Hs03023943_g1), GFAP (Hs00909233_m1), TUBB3 (Hs00801390_s1), PRNP (Hs01920617_s1) and PRNP_CDS (Hs04937277_s1).

Techniques: Produced, Western Blot, Bioprocessing, Staining, Expressing