negative control lentiviral shrna vectors  (Santa Cruz Biotechnology)

Journal: British Journal of Cancer

Article Title: Regulatory non-coding somatic mutations as drivers of neuroblastoma

doi: 10.1038/s41416-025-02939-0

Figure Lengend Snippet: Correlation between STAT3 and CTTNBP2 expression in a LAN-2 and b SH-SY5Y cells transfected with three pooled STAT3 siRNA. CTTNBP2 and STAT3 expression relative to their respective siScrambles (represented as single scramble bar in histogram), as measured by western blot (left) and qPCR (right) 24 h 48 h and 72 h post STAT3 silencing. β-Actin protein levels are used as loading control. c Luciferase reporter gene assay for CTTNBP2 CRE SNV1 carried out in LAN-2 and SH-SY5Y 72 h post STAT3 silencing. Luciferase activity of CTTNBP2 CRE SNV1 is normalized to that from cells transfected with wild-type construct and siScramble (scramble). d Allele-specific ChIP-qPCR conducted on plasmids carrying the wild-type or CTTNBP2 CRE SNV1 sequence upon transient transfection in LAN-2 and SH-SY5Y cell lines. Data is presented as fold-change of variant sequence upon comparing to wild-type sequence. All data shown are the mean ± standard deviation from three independent experiments each done in triplicate. Significant p -values obtained by two-tailed T-test are reported by * (* < 0.05; ** <0. 01; *** <0.001). Ref Reference, Alt Altered.

Article Snippet: STAT3, SIN3A, CTTNBP2 and MCF2L-specific siRNA-27 duplexes (pooled siRNA A-B-C) and SR30004-Universal Scrambled Negative Control siRNA (DS Scrambled Neg) duplex (Origene) were used at a concentration of 10 nM to transfect LAN-2, SH-SY5Y, CHP-212 and SK-N-BE(2) cells with X-tremeGENE siRNA Transfection Reagent (Roche).

Techniques: Expressing, Transfection, Western Blot, Control, Luciferase, Reporter Gene Assay, Activity Assay, Construct, ChIP-qPCR, Sequencing, Variant Assay, Standard Deviation, Two Tailed Test

Journal: British Journal of Cancer

Article Title: Regulatory non-coding somatic mutations as drivers of neuroblastoma

doi: 10.1038/s41416-025-02939-0

Figure Lengend Snippet: Correlation between SIN3A and CTTNBP2 expression in a LAN-2 and b SH-SY5Y transfected with three pooled SIN3A siRNA. CTTNBP2 and SIN3A expression relative to their respective siScrambles (reported as single scramble bar in histogram), as measured by western blot (left) and qPCR (right) 24 h, 48 h and 72 h post SIN3A silencing. β-Actin protein levels are used as loading control. c Luciferase reporter gene assay for CTTNBP2 CRE SNV2 carried out in LAN-2 and SH-SY5Y 72 h post SIN3A silencing. Luciferase activity of CTTNBP2 CRE SNV2 is normalized to that from cells transfected with wild-type construct and siScramble (scramble). d Allele-specific ChIP-qPCR conducted on plasmids carrying the wild-type or CTTNBP2 CRE SNV2 sequence upon transient transfection in LAN-2 and SH-SY5Y cell lines. Data is presented as fold-change of variant sequence upon comparing to wild-type sequence. All data shown are the mean ± standard deviation from three independent experiments, each done in triplicate. Significant p -values obtained by two-tailed T-test are reported by * (* < 0.05; ** <0.01; *** <0.001). Ns non-significant, p -value, Ref Reference, Alt Altered.

Article Snippet: STAT3, SIN3A, CTTNBP2 and MCF2L-specific siRNA-27 duplexes (pooled siRNA A-B-C) and SR30004-Universal Scrambled Negative Control siRNA (DS Scrambled Neg) duplex (Origene) were used at a concentration of 10 nM to transfect LAN-2, SH-SY5Y, CHP-212 and SK-N-BE(2) cells with X-tremeGENE siRNA Transfection Reagent (Roche).

Techniques: Expressing, Transfection, Western Blot, Control, Luciferase, Reporter Gene Assay, Activity Assay, Construct, ChIP-qPCR, Sequencing, Variant Assay, Standard Deviation, Two Tailed Test

Journal: British Journal of Cancer

Article Title: Regulatory non-coding somatic mutations as drivers of neuroblastoma

doi: 10.1038/s41416-025-02939-0

Figure Lengend Snippet: Correlation between STAT3 and MCF2L expression in a SK-N-BE(2) and b CHP-212 cells transfected with three pooled STAT3 siRNA. MCF2L and STAT3 expression relative to their respective siScramble (reported as a single scramble bar in histogram), as measured by western blot (left) and qPCR (right) 24 h, 48 h and 72 h post STAT3 silencing. β-Actin protein levels are used as loading control. c Luciferase reporter gene assay for MCF2L CRE SNV1 carried out in SK-N-BE(2) and CHP-212 72 h post STAT3 silencing. Luciferase activity of MCF2L CRE SNV1 is normalized to that from cells transfected with wild-type construct and siScramble (scramble). d Allele-specific ChIP-qPCR conducted on plasmids carrying the wild-type or MCF2L CRE SNV1 sequence upon transient transfection in SK-N-BE(2) and CHP-212 cell lines. Data is presented as fold-change of variant sequence upon comparing to wild-type sequence. All data shown are the mean ± standard deviation from three independent experiments, each done in triplicate. Significant p -values obtained by two-tailed T-test are reported by * *(* < 0.05; ** < 0.01; *** < 0.001). Ns non-significant p -value, Ref Reference, Alt Altered.

Article Snippet: STAT3, SIN3A, CTTNBP2 and MCF2L-specific siRNA-27 duplexes (pooled siRNA A-B-C) and SR30004-Universal Scrambled Negative Control siRNA (DS Scrambled Neg) duplex (Origene) were used at a concentration of 10 nM to transfect LAN-2, SH-SY5Y, CHP-212 and SK-N-BE(2) cells with X-tremeGENE siRNA Transfection Reagent (Roche).

Techniques: Expressing, Transfection, Western Blot, Control, Luciferase, Reporter Gene Assay, Activity Assay, Construct, ChIP-qPCR, Sequencing, Variant Assay, Standard Deviation, Two Tailed Test

Journal: British Journal of Cancer

Article Title: Regulatory non-coding somatic mutations as drivers of neuroblastoma

doi: 10.1038/s41416-025-02939-0

Figure Lengend Snippet: a CTTNBP2 (left) and MCF2L (right) expression by clinical features. From the top, risk group, MYCN amplification and INSS Stage. b In LAN-2 cells transfected with three pooled CTTNBP2 siRNA, CTTNBP2 expression relative to siScramble (scramble), as measured by western blot (left) and qPCR (right) 24 h, 48 h and 72 h post CTTNBP2 silencing. c SK-N-BE(2) cells transfected with three pooled MCF2L siRNA, MCF2L expression relative to siScramble (scramble), as measured by western blot (left) and qPCR (right) 24 h, 48 h and 72 h post MCF2L silencing. β-Actin protein levels are used as loading control. d Cell viability assays in LAN-2 (left) and SK-N-BE(2) (right) cells silenced for CTTNBP2 and MCF2L respectively, are shown as fold change compared to wild-type cells. e Invading cells number of LAN-2 (left) and SK-N-BE(2) (right) silenced for CTTNBP2 and MCF2L respectively, are shown as fold change compared to wild-type cells. Representative images (10×) of invasion assays in silenced cells are reported on the right. All data shown are the mean ± standard deviation from three independent qPCR experiments, each done in triplicate. Significant p -values obtained by two-tailed T-test are reported by * (* < 0.05; ** < 0.01; *** < 0.001).

Article Snippet: STAT3, SIN3A, CTTNBP2 and MCF2L-specific siRNA-27 duplexes (pooled siRNA A-B-C) and SR30004-Universal Scrambled Negative Control siRNA (DS Scrambled Neg) duplex (Origene) were used at a concentration of 10 nM to transfect LAN-2, SH-SY5Y, CHP-212 and SK-N-BE(2) cells with X-tremeGENE siRNA Transfection Reagent (Roche).

Techniques: Expressing, Amplification, Transfection, Western Blot, Control, Standard Deviation, Two Tailed Test