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sinrn1 pool  (siTools Biotech)
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siTools Biotech sinrn1 pool
Notch signaling and downstream targets in overexpression cell line and under knock-down. a qRT-PCR of mRNA expression levels of Hes1 and Hey1. Comparison of Notch downstream target expression between GFP and NRN1-GFP. Expression levels normalized to β-actin. GFP set to 1. b Protein expression of HES1. Analysis of protein levels of HES1 in GFP and NRN1-GFP cell lines through Western blot. Equal loading was controlled with GAPDH primary antibody. GFP set to 1. c Luciferase assay for analysis of Hes1 promotor activity. Comparison of Hes1 promotor activity between GFP and NRN1-GFP. Measurements normalized to transfection control pRL-TK. GFP set to 1. d Protein expression of N1ICD in nuclear extracts. Western blot of nuclear fractions comparing GFP and NRN1-GFP. Equal loading was controlled with LaminB2 primary antibody. GFP set to 1. e Protein expression of HES1 in nuclear extracts. Western blot of nuclear fractions comparing GFP and NRN1-GFP. Equal loading was controlled with LaminB2 primary antibody. GFP set to 1. f Analysis of mRNA levels of Hes1 under <t>siNRN1</t> knock-down. qRT-PCR comparing siCTR and siNRN1. Expression levels normalized to β-actin. siCTR set to 1. g Luciferase assay for analysis of Hes1 promotor activity under siNRN1 knock-down. Comparison of Hes1 promotor activity between siCTR and siNRN1. Measurements normalized to transfection control pRL-TK. siCTR set to 1. h Protein expression of HES1 under siNRN1 knock-down. Comparing protein levels of HES1 in siCTR and siNRN1 lysates through Western blot. Equal loading was controlled with β-actin primary antibody. siCTR set to 1. i Immunoprecipitation of NRN1-GFP lysates after transfection (control, N1ICD-V5). Lysates were pulled with IgG, V5 and Notch3 antibody. Western blot was probed with V5 primary antibody, Notch3 primary antibody and GFP primary antibody. Detected interaction complexes are denoted by black arrow. All graphs are displayed as mean ± SEM. Two groups were statistically analysed using unpaired Students t-test unless stated otherwise. * = p < 0.05, ns = p > 0.05
Sinrn1 Pool, supplied by siTools Biotech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Notch signaling and downstream targets in overexpression cell line and under knock-down. a qRT-PCR of mRNA expression levels of Hes1 and Hey1. Comparison of Notch downstream target expression between GFP and NRN1-GFP. Expression levels normalized to β-actin. GFP set to 1. b Protein expression of HES1. Analysis of protein levels of HES1 in GFP and NRN1-GFP cell lines through Western blot. Equal loading was controlled with GAPDH primary antibody. GFP set to 1. c Luciferase assay for analysis of Hes1 promotor activity. Comparison of Hes1 promotor activity between GFP and NRN1-GFP. Measurements normalized to transfection control pRL-TK. GFP set to 1. d Protein expression of N1ICD in nuclear extracts. Western blot of nuclear fractions comparing GFP and NRN1-GFP. Equal loading was controlled with LaminB2 primary antibody. GFP set to 1. e Protein expression of HES1 in nuclear extracts. Western blot of nuclear fractions comparing GFP and NRN1-GFP. Equal loading was controlled with LaminB2 primary antibody. GFP set to 1. f Analysis of mRNA levels of Hes1 under siNRN1 knock-down. qRT-PCR comparing siCTR and siNRN1. Expression levels normalized to β-actin. siCTR set to 1. g Luciferase assay for analysis of Hes1 promotor activity under siNRN1 knock-down. Comparison of Hes1 promotor activity between siCTR and siNRN1. Measurements normalized to transfection control pRL-TK. siCTR set to 1. h Protein expression of HES1 under siNRN1 knock-down. Comparing protein levels of HES1 in siCTR and siNRN1 lysates through Western blot. Equal loading was controlled with β-actin primary antibody. siCTR set to 1. i Immunoprecipitation of NRN1-GFP lysates after transfection (control, N1ICD-V5). Lysates were pulled with IgG, V5 and Notch3 antibody. Western blot was probed with V5 primary antibody, Notch3 primary antibody and GFP primary antibody. Detected interaction complexes are denoted by black arrow. All graphs are displayed as mean ± SEM. Two groups were statistically analysed using unpaired Students t-test unless stated otherwise. * = p < 0.05, ns = p > 0.05

Journal: Cell Communication and Signaling : CCS

Article Title: NRN1 interacts with Notch to increase oncogenic STAT3 signaling in melanoma

doi: 10.1186/s12964-024-01632-8

Figure Lengend Snippet: Notch signaling and downstream targets in overexpression cell line and under knock-down. a qRT-PCR of mRNA expression levels of Hes1 and Hey1. Comparison of Notch downstream target expression between GFP and NRN1-GFP. Expression levels normalized to β-actin. GFP set to 1. b Protein expression of HES1. Analysis of protein levels of HES1 in GFP and NRN1-GFP cell lines through Western blot. Equal loading was controlled with GAPDH primary antibody. GFP set to 1. c Luciferase assay for analysis of Hes1 promotor activity. Comparison of Hes1 promotor activity between GFP and NRN1-GFP. Measurements normalized to transfection control pRL-TK. GFP set to 1. d Protein expression of N1ICD in nuclear extracts. Western blot of nuclear fractions comparing GFP and NRN1-GFP. Equal loading was controlled with LaminB2 primary antibody. GFP set to 1. e Protein expression of HES1 in nuclear extracts. Western blot of nuclear fractions comparing GFP and NRN1-GFP. Equal loading was controlled with LaminB2 primary antibody. GFP set to 1. f Analysis of mRNA levels of Hes1 under siNRN1 knock-down. qRT-PCR comparing siCTR and siNRN1. Expression levels normalized to β-actin. siCTR set to 1. g Luciferase assay for analysis of Hes1 promotor activity under siNRN1 knock-down. Comparison of Hes1 promotor activity between siCTR and siNRN1. Measurements normalized to transfection control pRL-TK. siCTR set to 1. h Protein expression of HES1 under siNRN1 knock-down. Comparing protein levels of HES1 in siCTR and siNRN1 lysates through Western blot. Equal loading was controlled with β-actin primary antibody. siCTR set to 1. i Immunoprecipitation of NRN1-GFP lysates after transfection (control, N1ICD-V5). Lysates were pulled with IgG, V5 and Notch3 antibody. Western blot was probed with V5 primary antibody, Notch3 primary antibody and GFP primary antibody. Detected interaction complexes are denoted by black arrow. All graphs are displayed as mean ± SEM. Two groups were statistically analysed using unpaired Students t-test unless stated otherwise. * = p < 0.05, ns = p > 0.05

Article Snippet: For knockdowns using siNRN1 pool (Gene ID: 51299, siTools Biotech GmbH, Planegg, Germany) 1.5 x 10 5 cells were transfected with 5 nM siNRN1 pool while floating according to the Lipofectamine TM RNAiMAX (Thermofisher Scientific) protocol.

Techniques: Over Expression, Knockdown, Quantitative RT-PCR, Expressing, Comparison, Western Blot, Luciferase, Activity Assay, Transfection, Control, Immunoprecipitation

Expression of STAT3 targets under NRN1 overexpression. a qRT-PCR analysis of mRNA expression of Vegf A. Comparison of GFP and NRN1-GFP. Expression levels normalized to β-actin. GFP set to 1. b Vegf A promotor activity analysis. Luciferase-based assay comparing Vegf A promotor activity of GFP and NRN1-GFP cells. Measurements normalized to transfection control pRL-TK. GFP set to 1. c mRNA analysis of Mdr1 in GFP and NRN1-GFP cells through qRT-PCR. Expression levels normalized to β-actin. GFP set to 1. d Mdr1 promotor activity analysis using Luciferase-based assay comparing GFP and NRN1-GFP cells. Measurements normalized to transfection control pRL-TK. GFP set to 1. e Mdr1 promotor activity analysis using Luciferase-based assay comparing siCTR and siNRN1 knockdown cells. Measurements normalized to transfection control pRL-TK. siCTR set to 1. f cMet mRNA expression analysis using qRT-PCR of GFP and NRN1-GFP. GFP set to 1. All graphs are displayed as mean ± SEM. Two groups were statistically analysed using unpaired Students t-test unless stated otherwise. * = p < 0.05, ns = p > 0.05

Journal: Cell Communication and Signaling : CCS

Article Title: NRN1 interacts with Notch to increase oncogenic STAT3 signaling in melanoma

doi: 10.1186/s12964-024-01632-8

Figure Lengend Snippet: Expression of STAT3 targets under NRN1 overexpression. a qRT-PCR analysis of mRNA expression of Vegf A. Comparison of GFP and NRN1-GFP. Expression levels normalized to β-actin. GFP set to 1. b Vegf A promotor activity analysis. Luciferase-based assay comparing Vegf A promotor activity of GFP and NRN1-GFP cells. Measurements normalized to transfection control pRL-TK. GFP set to 1. c mRNA analysis of Mdr1 in GFP and NRN1-GFP cells through qRT-PCR. Expression levels normalized to β-actin. GFP set to 1. d Mdr1 promotor activity analysis using Luciferase-based assay comparing GFP and NRN1-GFP cells. Measurements normalized to transfection control pRL-TK. GFP set to 1. e Mdr1 promotor activity analysis using Luciferase-based assay comparing siCTR and siNRN1 knockdown cells. Measurements normalized to transfection control pRL-TK. siCTR set to 1. f cMet mRNA expression analysis using qRT-PCR of GFP and NRN1-GFP. GFP set to 1. All graphs are displayed as mean ± SEM. Two groups were statistically analysed using unpaired Students t-test unless stated otherwise. * = p < 0.05, ns = p > 0.05

Article Snippet: For knockdowns using siNRN1 pool (Gene ID: 51299, siTools Biotech GmbH, Planegg, Germany) 1.5 x 10 5 cells were transfected with 5 nM siNRN1 pool while floating according to the Lipofectamine TM RNAiMAX (Thermofisher Scientific) protocol.

Techniques: Expressing, Over Expression, Quantitative RT-PCR, Comparison, Activity Assay, Luciferase, Transfection, Control, Knockdown