Journal: Scientific Reports

Article Title: Novel variants in Krueppel like factor 1 that cause persistence of fetal hemoglobin in In(Lu) individuals

doi: 10.1038/s41598-021-97149-y

Figure Lengend Snippet: knockdown of endogenous KLF1 combined with re-expression of specific KLF1 variants show differential degrees of target gene expression rescue. Endogenous KLF1 was downregulated in primary human erythroblasts through transduction of short hairpin targeting 5’UTR of KLF1 mRNA. These cells were then selected by puromycin and transduced with wild type or mutant KLF1 and differentiated. (A) CD235 and CD71 expression levels do not overtly change upon knockdown or overexpression of wt or mutant KLF1. FCM plots show the differentiation state of erythroid cells at 72 h of differentiation. Cells in the GFP + plots (on the right axis of the plots) express mutant or wild type KLF1 and GFP- (left axis) do not ectopically express KLF1. Sh indicates co-expression with the shRNA against endogenous KLF1. (B) Lu/BCAM expression as depicted by FCM plots. Cells with lentiviral mediated expression of KLF1 wt or mutants are GFP + . (C) FCM plots shows the expression of CD44 in cells with or without an shRNA against endogenous KLF1 and with or without lentiviral induced expression of KLF1 wt or mutants (GFP +). (D–G) The expression (GM) of Blood Groups, CD235, CD44, Lu, andCD71 respectively, at 72 h of erythroid differentiation for each KLF1 mutant, wild type KLF1, in combination with shRNA against KLF1 or scrambled control.

Article Snippet: The ORF from human KLF1 was amplified by PCR using primers with specific restriction enzymes within the oligo’s and cloned into the lentiviral vector PAD5 (pRRLSIN.PPT.hCMV.2A-GFPpre ), which was cut using XbaI (NEB, R0145S) and EcoRI (NEB, R0101S).

Techniques: Expressing, Transduction, Mutagenesis, Over Expression, shRNA

Journal: The Journal of Infectious Diseases

Article Title: Cytomegalovirus DNAemia in Hospitalized Adults With SARS-CoV-2 Infection Requiring Supplemental Oxygen: Virologic and Clinical Characteristics and Association With Outcomes

doi: 10.1093/infdis/jiaf649

Figure Lengend Snippet: Multivariate analysis of maximum SARS-CoV-2 viral load between baseline and day 15 ( A ) and between day 15 and 29 ( B ) among day 15 survivors. Mean differences refer to cycle threshold values obtained by polymerase chain reaction. Abbreviations: CI, confidence interval; CMV, cytomegalovirus; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2.

Article Snippet: In CMV-seropositive trial participants, baseline and longitudinal plasma samples were tested by quantitative CMV plasma polymerase chain reaction (PCR; Abbott RealTime CMV assay, Abbott Laboratories, Chicago, IL, USA).

Techniques: Polymerase Chain Reaction