Journal: Wellcome Open Research

Article Title: ZMYM2 inhibits NANOG-mediated reprogramming

doi: 10.12688/wellcomeopenres.15250.1

Figure Lengend Snippet: Probes used for qPCR.

Article Snippet: Nr0b1 , Mm00431729_m1.

Techniques:

Journal: PLoS ONE

Article Title: Inhibition of β-catenin dependent WNT signalling upregulates the transcriptional repressor NR0B1 and downregulates markers of an A9 phenotype in human embryonic stem cell-derived dopaminergic neurons: Implications for Parkinson’s disease

doi: 10.1371/journal.pone.0261730

Figure Lengend Snippet: Panels (a) and (b) respectively show the effects of the WNT modulators and NEUROD1 overexpression upon NR0B1 . Panels (c) and (d) show effects upon NR0B2 expression. Results are expressed as mean ± SEM of three (biological) replicate experiments. Analysis of panel (a) was via one-way ANOVA with post-hoc Dunnett’s test. For panels (b) and (d) analysis was via Student’s t-tests (at each time point as in ). For panel (c) analysis was via a non-parametric Kruskal-Wallis test with post-hoc Dunn’s test to accommodate undetectable levels of NR0B2 in two out of three samples, both undetectable values were assigned to 0.0028 (equal to the lowest value detected).

Article Snippet: At this time, cultures were transfected with NEUROD1 (SC118625, Origene), NR0B1 (SC319624, Origene), NR0B2 (SC122920, Origene) or blank plasmids (PCMV6XL5, Origene) according to manufacturer’s instructions.

Techniques: Over Expression, Expressing

Journal: PLoS ONE

Article Title: Inhibition of β-catenin dependent WNT signalling upregulates the transcriptional repressor NR0B1 and downregulates markers of an A9 phenotype in human embryonic stem cell-derived dopaminergic neurons: Implications for Parkinson’s disease

doi: 10.1371/journal.pone.0261730

Figure Lengend Snippet: Expression patterns following transfection with NR0B1 (green), NR0B2 (red) or blank (blue) plasmids show that NR0B1 transfection increases both NR0B1 and NR0B2 and impacts expression of NEUROD1 , TH , KCNJ6 , LMX1A and TUBB3 . Control wells underwent transfection with an empty plasmid. Results are expressed as mean ± SEM of three (biological) replicate experiments. Analysis was performed with One-way ANOVA followed by post-hoc Dunnett’s test where each post transfection timepoint represented a repeated measure (t = 0 was not included in analyses). * and ** indicate P<0.05 and P<0.01, respectively. shows increased NR0B1 and NR0B2 immunolabelling up to six days after transfection.

Article Snippet: At this time, cultures were transfected with NEUROD1 (SC118625, Origene), NR0B1 (SC319624, Origene), NR0B2 (SC122920, Origene) or blank plasmids (PCMV6XL5, Origene) according to manufacturer’s instructions.

Techniques: Expressing, Transfection, Control, Plasmid Preparation

Journal: Nature Communications

Article Title: Dax1 modulates ERα-dependent hypothalamic estrogen sensing in female mice

doi: 10.1038/s41467-023-38618-y

Figure Lengend Snippet: A qPCR screening of 84 nuclear receptor co-regulators in the mouse arcuate and AVPV hypothalamus under estrogen stimulation ( n = 4). B Confirmation of Dax1 expression in the arcuate and AVPV hypothalamus using biological replicates under estrogen stimulation ( n = 4) * p < 0.05 by t-test error bars represent SEM. C Detection of the DAX1 protein (red) by immunofluorescence in the whole arcuate and in Kiss1 -neurons (green) in intact Kiss1 -Cre-GFP mice in diestrous ( n = 3) * p < 0.05 by t-test Error bars represent SEM. Scale bar 50 µm (left), 20 µm (right). D DAX1 and POMC in human hypothalamus ( n = 3). Error bars represent SEM. E expression of POMC and DAX1 in human arcuate nucleus ( n = 3) Error bars represent SEM. F CRISPR-activation of the Golt1a promotor in C2C12 cells transfected with mERα or/and mDax1. Representative data from an experiment conducted twice ( n = 4 wells per group) * p < 0.05 compared to Golt1a by Holm-Sidak multiple t-tests. Error bars represent SD. Source data are provided as a Source Data file.

Article Snippet: The RNA was then used to generate to cDNA via MMLV reverse transcriptase (Promega). qPCR was performed on human samples using TaqMan probes specific for NR0B1 (Hs05033649_g1), POMC (Hs01596743_m1) and GAPDH (Hs02786624_g1) and Taqman Universal PCR Master Mix (ThermoFisher) on an Applied Biosystems Quantstudio 7 qPCR instrument (ThermoFisher).

Techniques: Expressing, Immunofluorescence, CRISPR, Activation Assay, Transfection

Journal: Nature Communications

Article Title: Dax1 modulates ERα-dependent hypothalamic estrogen sensing in female mice

doi: 10.1038/s41467-023-38618-y

Figure Lengend Snippet: A quantification of Cre -mediated deletion of DAX1-protein in arcuate Kiss1 -neurons ( n = 3 mice). B Expression of Kiss1 , Tac2 , and Pdyn in the arcuate nucleus of intact mice at 9am on the first day of diestrous ( n = 8 Dax1 tm1 , n = 12 Dax1 th1(Kiss1) * p < 0.05. C Cycle length ( n = 8), Offipring/3-months ( n = 8 Dax1 tm1 , n = 10 Dax1 th1(Kiss1) ) * p < 0.05 by t-test, Tertiary follicles per ovary ( n = 5). ( D ) Plasma FSH levels at 9am on the first day of diestrous ( n = 7). E Representative LH profiles starting at 9am on the first day of diestrous. Peak LH ( n = 8) an inter-pulse interval ( n = 5 Dax1 tm1 , n = 4 Dax1 th1(Kiss1) ). F Kiss1 -expression in the AVPV and arcuate following estrogen stimulation ( n = 8) * p < 0.05 compared to vehicle 2-way ANOVA with Tukey’s multiple test correction. G Schematic representation of ERα function during negative feedback in arcuate Kiss1 -neurons (Created with BioRender.com). Error bars represent SEM. Source data are provided as a Source Data file.

Article Snippet: The RNA was then used to generate to cDNA via MMLV reverse transcriptase (Promega). qPCR was performed on human samples using TaqMan probes specific for NR0B1 (Hs05033649_g1), POMC (Hs01596743_m1) and GAPDH (Hs02786624_g1) and Taqman Universal PCR Master Mix (ThermoFisher) on an Applied Biosystems Quantstudio 7 qPCR instrument (ThermoFisher).

Techniques: Expressing, Clinical Proteomics