shrnas against sox2  (Vector Biolabs)

Journal: Neuro-Oncology

Article Title: SOX2 commands LIM homeobox transcription factors in choroid plexus development and tumorigenesis

doi: 10.1093/neuonc/noaf085

Figure Lengend Snippet: SnRNA-seq reveals cellular diversity and dynamic differentiation in NOTCH-driven CP tumors characterized by a glial progenitor-like signature. (A) Major cell types of a NOTCH-driven CPP from an adult Lcre;NICD1 mouse. Uniform Manifold Approximation and Projection (UMAP) of 6428 single-nucleus profiles, colored by post hoc annotated cell type; also see . (B) UMAP showing mesenchymal, epithelial, and NOTCH-activated profiles from snRNA-seq in NOTCH-driven CPP, colored by expression of Wnt5a (mesenchymal marker, left ), Hes1 (NOTCH pathway target, middle ), and Otx2 (CP epithelial marker, right ); also see . (C) GO analysis of differentially expressed genes in the epithelial-like tumor cell compartment in NOTCH-driven CPP. (D) The expression of markers for glial progenitors in the rhombic lip in NOTCH-driven CPP. UMAP shows 6428 single-nucleus profiles from snRNA-seq in NOTCH-driven CPP, colored by expression of genes associated with glial progenitors in the rhombic lip in the hindbrain ( Rspo1 , Zfp423 , Zic3 , Msx1 , Sox2 , and Slc1a3 ). (E) Western blot analysis of MSX1 expression in the CP of wild-type mice, and NOTCH-driven CPP in Lcre;NICD1 animals ( n = 3 per genotype, mean ± SEM, 2-tailed unpaired t test, **** P < 0.0001). Data were generated from 2 independent experiments. (F) RT-qPCR analysis of Rspo1 and Gdf7 mRNA levels in NOTCH-driven CPP and wild-type CP ( n = 3 per time point per genotype, mean ± SEM, 2-tailed unpaired t test, **** P < 0.0001). Three independent experiments were conducted. (G) UMAP showing single-nucleus profiles in the epithelial-like compartment in (A) colored by subgroups; also see . (H) Cell trajectory analysis of single-nucleus profiles of subgroups of the epithelial-like compartment, and the glia-like compartment in NOTCH-driven CPP. (I) Violin plots for the expression of Rspo1 , Zfp423 , Msx1 , and Zic4 in subgroups of the epithelial-like compartment in NOTCH-driven CPP.

Article Snippet: Viruses expressing HA-tagged LMX1A or LMX1B (269200540200, 269210540200, Applied Biological Materials Inc.), and viruses expressing SOX2, shRNAs against Sox2 (shADV-272885) or Lmx1a (shADV-263539, all from Vector Biolabs) were amplified and purified from AD-293 cells (Agilent Technologies).

Techniques: Expressing, Marker, Western Blot, Generated, Quantitative RT-PCR

Journal: Neuro-Oncology

Article Title: SOX2 commands LIM homeobox transcription factors in choroid plexus development and tumorigenesis

doi: 10.1093/neuonc/noaf085

Figure Lengend Snippet: Increased SOX2 expression in CP tumors in humans and mice. (A) Immunohistochemistry of SOX2 is shown in the upper rhombic lip/roof plate (red dotted lines) and hindbrain CP (arrow) in wild-type mice, NOTCH-driven CPP and CPC (arrowheads) in Lcre;NICD1 and Lcre;Ptch cko ;NICD1 animals at embryonic (E) day 14.5 (E14.5). Scale bar, 50 µm. Black dotted line marks the border of the ventricle with SOX2-expressing ependymal cells. (B) Immunofluorescence of SOX2 and OTX2 is shown in Rb1/Trp53 -deficient CPC in adult Lcre;p53 cko ;Rb cko mice. Nuclei are labeled with DAPI. Scale bar, 50 µm. (C) RT-qPCR analysis of Sox2 expression in wild-type CP, NOTCH-driven CPP, and Rb1/Trp53 -deficient CPC ( n = 5 per tissue type, mean ± SEM, 1-way ANOVA, **** P < 0.0001). (D) RNAscope of Sox2 and Myc expression in hindbrain CP (black arrow) in adult wild-type mice, and NOTCH-driven CPP (arrowhead) in adult Lcre;NICD1 animals. The dotted line marks ventricular walls with ependymal cells (red arrow). Scale bar, 50 µm. (E) Immunofluorescence of SOX2 and ARL13B is shown in NOTCH-driven CPP in adult Lcre;NICD1 animals. DAPI labels nuclei. Scale bar, 20 µm. (F) Immunofluorescence of SOX2 is shown in NOTCH-driven CP tumor cells treated with DMSO or RIN-1 (10 µM). DAPI labels nuclei. Scale bar, 50 µm. Quantification of fluorescence intensity is shown ( n = 333 [DMSO]; n = 101 [RIN-1], mean ± SEM, 2-tailed unpaired t test, **** P < 0.0001). (G) Immunohistochemistry of SOX2 in human CP tumors are shown. Scale bar, 50 µm. (CPC: n = 14; CPP and atypical CPP: n = 26). All results were obtained from 3 independent experiments. (H) RT-qPCR analysis of gene expression in human CP organoids and CP tumors ( n = 6 [CP organoid], n = 9 [CPP], n = 13 [CPC], mean ± SEM, 1-way ANOVA, * P < 0.05; ** P < 0.01; *** P < 0.001; NS, nonsignificant). (I) CosMx analysis of the expression of NOTCH1 , NOTCH2 , NOTCH3 , and SOX2 in a human CPC sample. Boxed region is shown in higher magnification on the right. (J) RNAscope studies of SOX2 and HES5 expression in a human CPP. Scale bar, 200 µm.

Article Snippet: Viruses expressing HA-tagged LMX1A or LMX1B (269200540200, 269210540200, Applied Biological Materials Inc.), and viruses expressing SOX2, shRNAs against Sox2 (shADV-272885) or Lmx1a (shADV-263539, all from Vector Biolabs) were amplified and purified from AD-293 cells (Agilent Technologies).

Techniques: Expressing, Immunohistochemistry, Immunofluorescence, Labeling, Quantitative RT-PCR, RNAscope, Fluorescence, Gene Expression

Journal: Neuro-Oncology

Article Title: SOX2 commands LIM homeobox transcription factors in choroid plexus development and tumorigenesis

doi: 10.1093/neuonc/noaf085

Figure Lengend Snippet: SOX2 is essential for the glial progenitor-like signature and NOTCH-driven CP tumor development. (A) Immunofluorescence of Ki-67 is shown in NOTCH-driven CPP cells treated with control scrambled siRNAs or siRNAs against Sox2 at different concentrations. DAPI labels nuclei. Scale bar, 50 µm. Quantification of Ki-67 expression is shown ( n = 3 per group, mean ± SEM, 1-way ANOVA, **** P < 0.0001). Data were obtained from 3 independent experiments; see also . (B) Bisected brain hemispheres and hematoxylin and eosin (H&E) staining of the hindbrain CP in adult wild-type mice, and CPP in adult Lcre;NICD1 and Lcre;NICD1;Sox2 cko animals. Red arrows point to wild-type CP, arrowheads point to CPP, black arrows point to SOX2-deficient CPP. Scale bar, 50 µm. (C) Immunofluorescence of Ki-67 and GFP is shown in CPP in Lcre;NICD1 and Lcre;NICD1;Sox2 cko animals at day E13.5. Nuclei are labeled with DAPI. Scale bar, 50 µm. Quantification of Ki-67 expression is shown ( n = 3 per group, mean ± SEM, 2-tailed unpaired t test, **** P < 0.0001). Results were obtained from 3 independent experiments; also see . (D) Principal component analysis (PCA) of CP in wild-type mice, CPP in Lcre;NICD1 mice, and Sox2 -deficient CPP in Lcre;NICD1;Sox2 cko animals; also see . (E) Gene set enrichment analysis (GSEA) of the effect of Sox2 loss on NOTCH-driven CP tumors. Pathways regulating pluripotency of the stem cells are shown as an example; also see . NES, normalized enrichment score. FDR, false discovery rate. (F) RT-qPCR analysis of gene expression in CP in wild-type mice, and CPP from Lcre;NICD1 and Lcre;NICD1;Sox2 cko animals ( n = 8 [CP], n = 6 [CPP], mean ± SEM, 1-way ANOVA, ** P < 0.01; *** P < 0.001; **** P < 0.0001; NS, nonsignificant); also see .

Article Snippet: Viruses expressing HA-tagged LMX1A or LMX1B (269200540200, 269210540200, Applied Biological Materials Inc.), and viruses expressing SOX2, shRNAs against Sox2 (shADV-272885) or Lmx1a (shADV-263539, all from Vector Biolabs) were amplified and purified from AD-293 cells (Agilent Technologies).

Techniques: Immunofluorescence, Control, Expressing, Staining, Labeling, Quantitative RT-PCR, Gene Expression

Journal: Neuro-Oncology

Article Title: SOX2 commands LIM homeobox transcription factors in choroid plexus development and tumorigenesis

doi: 10.1093/neuonc/noaf085

Figure Lengend Snippet: SOX2 transcriptional targets in CP tumors are enriched in markers and transcriptional regulators of progenitors in the rhombic lip. (A) Pie chart illustrating the distribution of SOX2-binding sites in relation to genes in NOTCH-driven CPP in Lcre;NICD1 animals; also see . (B) Heatmap of tag densities of SOX2 (left) or H3K23Ac (right) ChIP-seq signals at all of the binding regions identified in ChIP-seq experiments. In each heat map the tag density is plotted for 10 Kb at either side of its binding peak summit. (C) Comparison of SOX2 and H3K27Ac signals generated from ChIP-seq fragment counts in the 20 Kb genomic regions surrounding SOX2 peaks in NOTCH-driven CPP. (D) Logos for the motif enriched in SOX2-binding sequences identified by motif analysis in NOTCH-driven CPP; also see . TF: transcription factor; FDR: false discovery rate. (E) Venn diagram shows the overlap of SOX2-associated genes and differentially expressed genes in NOTCH-driven CPPs at days P0 and P21, respectively. (F) GO analysis of candidate SOX2 transcriptional targets in NOTCH-driven CPP. (G) Venn diagram shows the overlap of SOX2 candidate transcriptional targets in (E) and significantly downregulated genes in Sox2 -deficient tumors. (H) Hierarchical clustering of the expression of 52 candidate SOX2 transcriptional targets identified in (G, FDR < 0.05) in wild-type CP, and Sox2 -wild-type or Sox2 -deficient NOTCH-driven CPP. Lmx1b and Hes5 on the heatmap are marked by arrows. (I) The peak density plot of fragment counts is shown in genomic regions that encompass Lmx1b , Lmx1a , Hes5 , and Zic4 , and bound by SOX2 and H3K27Ac, respectively. Genes are labeled in black with sequence in a single exon as a rectangle; also see . (J) RT-qPCR analysis of gene expression in CP in wild-type mice, and CPP from Lcre;NICD1 and Lcre;NICD1;Sox2 cko animals ( n = 8 [CP], n = 6 [CPP], mean ± SEM, 1-way ANOVA, *** P < 0.001; **** P < 0.0001; NS, nonsignificant).

Article Snippet: Viruses expressing HA-tagged LMX1A or LMX1B (269200540200, 269210540200, Applied Biological Materials Inc.), and viruses expressing SOX2, shRNAs against Sox2 (shADV-272885) or Lmx1a (shADV-263539, all from Vector Biolabs) were amplified and purified from AD-293 cells (Agilent Technologies).

Techniques: Binding Assay, ChIP-sequencing, Comparison, Generated, Expressing, Labeling, Sequencing, Quantitative RT-PCR, Gene Expression

Journal: Neuro-Oncology

Article Title: SOX2 commands LIM homeobox transcription factors in choroid plexus development and tumorigenesis

doi: 10.1093/neuonc/noaf085

Figure Lengend Snippet: SOX2 regulates transcription factors LMX1A and LMX1B in NOTCH-driven CP tumors. (A) UMAP of 6428 single-nucleus profiles from a NOTCH-driven CPP colored by Lmx1a and Lmx1b expression, respectively. (B) t-distributed stochastic neighbor embedding (t-SNE) plot shows the annotated scATAC-seq profiles of different cell populations in NOTCH-driven CPP. Different subclusters of cells are marked by different colors. (C) Violin plots show the activity of different genes in each subcluster of cells. (D) Western blot analysis of the expression of SOX2, LMX1A, and LMX1B in CP in wild-type mice, and NOTCH-driven CPP in Lcre;NICD1 animals ( n = 3 per group, mean ± SEM, 2-tailed unpaired t test, ** P < 0.01; *** P < 0.001). Three independent experiments were conducted; also see . (E, F) Immunofluorescence of LMX1A (E) and LMX1B (F) is shown in Rb1/Trp53 -deficient CPC cells infected with viruses expressing SOX2. SOX2 labels infected cells. Scale bars, 50 µm. Three independent experiments were conducted. (G) Immunofluorescence of LMX1A is shown in TP53 -deficient human CPC cells infected with viruses expressing SOX2. SOX2 labels infected cells. Scale bar, 50 µm. Three independent experiments were conducted. (H, I) Immunofluorescence of LMX1A (H) LMX1B (I) is shown in CPP in Lcre;NICD1 and Lcre;NICD1;Sox2 cko animals at postnatal (P) day 7 (P7). Arrowheads point to tumor cells, arrows point to SOX2-deficient tumor cells. DAPI labels nuclei. Scale bars, 50 µm. Fluorescence intensity is quantified ( n = 1101 [LMX1A], n = 754 [LMX1B] for NOTCH-driven CPP cells; n = 457 [LMX1A], n = 563 [LMX1B] for Sox2 -deficient tumor cells; mean ± SEM, 2-tailed unpaired t test, **** P < 0.0001). The experiments were repeated 3 times independently; also see and . (J) RT-qPCR analysis of the expression of Lmx1a and Lmx1b in CP in wild-type mice, and CPP from Lcre;NICD1 and Lcre;NICD1;Sox2 cko animals ( n = 8 [CP], n = 6 [CPP], mean ± SEM, 1-way ANOVA, *** P < 0.001; **** P < 0.0001; NS, nonsignificant). Results were obtained from 1 experiment. (K) t-SNE plots show that motifs of LMX1A and LMX1B are enriched in tumor cell subclusters in NOTCH-driven CPP. Colors represent average gene activity score of cells in each subcluster. Dark red means high gene activity score, blue means low gene activity score.

Article Snippet: Viruses expressing HA-tagged LMX1A or LMX1B (269200540200, 269210540200, Applied Biological Materials Inc.), and viruses expressing SOX2, shRNAs against Sox2 (shADV-272885) or Lmx1a (shADV-263539, all from Vector Biolabs) were amplified and purified from AD-293 cells (Agilent Technologies).

Techniques: Expressing, Activity Assay, Western Blot, Immunofluorescence, Infection, Fluorescence, Quantitative RT-PCR

Journal: Neuro-Oncology

Article Title: SOX2 commands LIM homeobox transcription factors in choroid plexus development and tumorigenesis

doi: 10.1093/neuonc/noaf085

Figure Lengend Snippet: LMX1A and LMX1B mediate SOX2 functions to support tumor cell proliferation. (A) Immunofluorescence of Ki-67 is shown in NOTCH-driven CPP treated with control scrambled siRNAs or siRNAs against Lmx1a and/or Lmx1b (40 nM). DAPI labels nuclei. Scale bar, 50 µm. Quantification of Ki-67 expression is shown ( n = 3 per group, mean ± SEM, 1-way ANOVA, ** P < 0.01). The experiments were repeated 3 times independently; also see . (B) Immunofluorescence of Ki-67 is shown in NOTCH-driven CPP treated with control scrambled siRNAs or siRNAs against Sox2 (40 nM), and infected with viruses expressing HA-tagged Lmx1a , Lmx1b , or control viruses. DAPI labels nuclei. Scale bar, 50 µm. Quantification of Ki-67 expression is shown ( n = 3 per group, mean ± SEM, 1-way ANOVA, **** P < 0.0001; NS, nonsignificant). Three independent experiments were conducted; also see . (C) Immunofluorescence of LMX1B is shown in NOTCH-driven CPP cells treated with DMSO or RIN-1 (10 µM). DAPI labels nuclei. Scale bar, 50 µm. Quantification of fluorescence intensity is shown ( n = 233 [DMSO]; n = 201 [RIN-1], mean ± SEM, 2-tailed unpaired t test, **** P < 0.0001). (D) RT-qPCR analysis of the expression of SOX2 , LMX1A , and LMX1B in human CP organoids and CP tumors ( n = 6 [CP organoid], n = 9 [CPP], n = 13 [CPC]; mean ± SEM, NS, nonsignificant). The experiments were repeated 1 time independently. (E) RNAscope studies of LMX1A and SOX2 expression in human CP tumors. Scale bar, 200 µm. (F) Immunofluorescence of LMX1A in human CP tumor samples is shown. DAPI labels nuclei. Scale bar, 50 µm. Three independent experiments were conducted. (G) RNAscope studies of LMX1A and HES5 expression in a human CPP. Scale bar, 200 µm.

Article Snippet: Viruses expressing HA-tagged LMX1A or LMX1B (269200540200, 269210540200, Applied Biological Materials Inc.), and viruses expressing SOX2, shRNAs against Sox2 (shADV-272885) or Lmx1a (shADV-263539, all from Vector Biolabs) were amplified and purified from AD-293 cells (Agilent Technologies).

Techniques: Immunofluorescence, Control, Expressing, Infection, Fluorescence, Quantitative RT-PCR, RNAscope

Journal: Nature communications

Article Title: DNMT3B supports meso-endoderm differentiation from mouse embryonic stem cells.

doi: 10.1038/s41467-023-35938-x

Figure Lengend Snippet: Fig. 3 | Rescue of the meso-endoderm phenotype in Dnmt3b−/−cells. a Western blot analysis of DNMT3B expression in 3BKO (two independent clones) EpiLC derived from ESCs transfected with Empty vector (control) or Avitag-Dnmt3b (Rescued). WT cells and β-actin serve as control. Representative of two indepen- dent experiments. Uncropped gels are provided in Supplementary Fig. 11. b Trends of gene expression dynamics for all the DEGs (activated at the top, repressed at the bottom) between 3BKO + empty (3BKO) (orange) and 3BKO + 3B (rescued) (purple) during differentiation from EpiLC to ME24 and ME48h. Genes are grouped as early,

Article Snippet: Sox2 silencing in Dnmt3b mutant cells Custom shRNAs against Sox2 were constructed using the TRC hairpin design tool (http://www.broadinstitute.org/rnai/public/seq/search), and designed to target the following sequences: 5′-ACCAATCCCATCCAAATTAAC-3′ (shRNA1) 5′-GCACAGTTTGAGATAAATAAA-3′ (shRNA2) Hairpins were cloned into pLKO.1 vector (Addgene plasmid_#10878) and each construct was verified by sequencing. a 0 1 D im .2 Clone B126 B77 WT MFA [ RNA-seq + WGBS ] b 3BKO upregulated genes [expression] d GO: Biological Process [top 10] ea rly m id la te microtubule bundle formation axoneme assembly male meiotic nuclear division negative reg. of cell morphogenesis involved in differentiation cilium organization intrinsic apoptotic signaling pathway in response to DNA damage pattern specification process telencephalon cell migration negative reg. of neuron projection development cilium movement import into cell reg. of cation channel activity calcium ion transmembrane transport positive reg.of transporter activity positive reg.of ion transport reg. of transporter activity axonogenesis hindbrain morphogenesis central nervous system neuron differentiation cellular response to virus dendrite development negative reg.of nervous system development reg.of cell morphogenesis involved in differentiation regionalization urogenital system development 0 2 4 6 c m id la te Gli2 Sox2 Nnat Sox1 Id2 Olig3 WGBS Z score (logRPKM) Regulatory Region D iff er en tia lly in du ce d ge ne s Methylation [%] 0 50 100 Promoters SuperEnhancers TypicalEnhancers WT 3BKOWT 3BKO Differentiation [direct targets] Stage-Genotype 1# 2# 1# 2#1# 2#1# 2# B7 7 B1 26 B1 26 B7 7 B1 26 B7 7 B1 26 B7 71# 2# 1# 2#1# 2#1# 2# 1# 2# 1# 2#1# 2#1# 2# 1# 2# 1# 2#1# 2#1# 2# B126 B77 B126 B77 B126 B126B77 B77 ESC EpiLC ME 24h ME 48h ESC EpiLC ME 24h ME 48h ESC EpiLC ME 24h ME 48h ESC EpiLC ME24h ME48h ESC-WT EpiLC-WT ME24h-WT ME48h-WT ESC-3BKO EpiLC-3BKO ME24h-3BKO ME48h-3BKO Stage-Genotype ESC-WT EpiLC-WT ME24h-WT ME48h-WT ESC-3BKO EpiLC-3BKO ME24h-3BKO ME48h-3BKO 3BKO-B126 3BKO-B77 WT#1 WT#2 A ve ra ge e xp re ss io n [s ca le d R P K M ] A ve ra ge m et hy la tio n [% ] W G B S early mid late ES C Ep iLC M E2 4h M E4 8h ES C Ep iLC M E2 4h M E4 8h ES C Ep iLC M E2 4h M E4 8h 25 50 75 Nature Communications | (2023) 14:367 10 Oligonucleotide sequences for shRNA cloning are reported in Supplementary Data 8.

Techniques: Western Blot, Expressing, Clone Assay, Derivative Assay, Transfection, Plasmid Preparation, Control, Gene Expression

Journal: Journal of Southern Medical University

Article Title: SOX2-OT/SOX2 axis regulates lung cancer H520 cell migration via Gli1-mediated epithelial-mesenchymal transition

doi: 10.12122/j.issn.1673-4254.2022.10.01

Figure Lengend Snippet: Sequences of the primers for qRT-PCR

Article Snippet: Short hairpin RNA (shRNA) plasmids against SOX2-OT were purchased from Genechem (Shanghai, China).

Techniques:

Journal: Journal of Southern Medical University

Article Title: SOX2-OT/SOX2 axis regulates lung cancer H520 cell migration via Gli1-mediated epithelial-mesenchymal transition

doi: 10.12122/j.issn.1673-4254.2022.10.01

Figure Lengend Snippet: Expression level of SOX2-OT mRNA in lung squamous cell carcinoma (LUSC) specimens and in different lung cancer cell lines. A: Scatter plot generated by GEPIA for comparing transcription levels of SOX2-OT between 486 LUSC and 338 adjacent normal tissues. B: qRT-PCR analysis showing aberrantly higher SOX2-OT expression in H520 cells than in the other cell lines. C: SOX2-OT transcription level in H520 cells after transfection with two shRNAs (#4 and #5) for SOX2-OT knockdown and the empty vector (SHV). β-actin was used as the internal control. All data are presented as Mean±SD if not specified otherwise. *P < 0.05, **P < 0.01.

Article Snippet: Short hairpin RNA (shRNA) plasmids against SOX2-OT were purchased from Genechem (Shanghai, China).

Techniques: Expressing, Generated, Quantitative RT-PCR, Transfection, Plasmid Preparation

Journal: Journal of Southern Medical University

Article Title: SOX2-OT/SOX2 axis regulates lung cancer H520 cell migration via Gli1-mediated epithelial-mesenchymal transition

doi: 10.12122/j.issn.1673-4254.2022.10.01

Figure Lengend Snippet: Effect of SOX2-OT knockdown on invasive capacity of H520 cells. SOX2-OT knockdown significantly inhibits metastatic capacity of H520 cells as shown by wound healing assay (A, B; original magnification: ×100) and attenuates invasiveness of the cells as shown by Transwell assay (C, D; ×200). **P < 0.01.

Article Snippet: Short hairpin RNA (shRNA) plasmids against SOX2-OT were purchased from Genechem (Shanghai, China).

Techniques: Wound Healing Assay, Transwell Assay

Journal: Journal of Southern Medical University

Article Title: SOX2-OT/SOX2 axis regulates lung cancer H520 cell migration via Gli1-mediated epithelial-mesenchymal transition

doi: 10.12122/j.issn.1673-4254.2022.10.01

Figure Lengend Snippet: Effect of modification of SOX2-OT expression on Gli1 expression in H520 cells. SOX2-OT knockdown in H520 cells down-regulates expressions of Gli1 mRNA and protein detected with qRT-PCR (A) and Western blotting (B, C). Transfection with Gli1-overexpressing plasmid (pcDNA3.1-Gli1) counteracts the inhibitory effect of SOX2-OT knockdown shown by qRT-PCR (D) and Western blotting (E, F). Cells co-transfected with SHV and pcDNA3.1 backbone served as the control. **P < 0.01.

Article Snippet: Short hairpin RNA (shRNA) plasmids against SOX2-OT were purchased from Genechem (Shanghai, China).

Techniques: Modification, Expressing, Quantitative RT-PCR, Western Blot, Transfection, Plasmid Preparation

Journal: Journal of Southern Medical University

Article Title: SOX2-OT/SOX2 axis regulates lung cancer H520 cell migration via Gli1-mediated epithelial-mesenchymal transition

doi: 10.12122/j.issn.1673-4254.2022.10.01

Figure Lengend Snippet: Changes of expressions of EMT-related genes inH520 cells with SOX2-OT knockdown detected by qRT-PCR (A) and Western blotting (B, C) with β-actin and α- tubulin as the inner control, respectively. *P < 0.05, **P < 0.01.

Article Snippet: Short hairpin RNA (shRNA) plasmids against SOX2-OT were purchased from Genechem (Shanghai, China).

Techniques: Quantitative RT-PCR, Western Blot

Journal: Journal of Southern Medical University

Article Title: SOX2-OT/SOX2 axis regulates lung cancer H520 cell migration via Gli1-mediated epithelial-mesenchymal transition

doi: 10.12122/j.issn.1673-4254.2022.10.01

Figure Lengend Snippet: Gli1 overexpression attenuates the inhibitory effect of SOX2-OT knockdown on cell invasion and EMT-related phenotype in H520 cells. A, B: Wound healing assay of H520 cells transfected with both shRNA against SOX2-OT (#4) and pcDNA 3.1- Gli1 (× 100). C, D: Transwell assay of H520 cells of H520 cells transfected with both shRNA against SOX2-OT (#4) and pcDNA 3.1-Gli1 (×200). E: mRNA expression levels of vimentin, Snail and E-cadherin in H520 cells with different treatments detected with qRT-PCR. **P < 0.01.

Article Snippet: Short hairpin RNA (shRNA) plasmids against SOX2-OT were purchased from Genechem (Shanghai, China).

Techniques: Over Expression, Wound Healing Assay, Transfection, shRNA, Transwell Assay, Expressing, Quantitative RT-PCR

Journal: Journal of Southern Medical University

Article Title: SOX2-OT/SOX2 axis regulates lung cancer H520 cell migration via Gli1-mediated epithelial-mesenchymal transition

doi: 10.12122/j.issn.1673-4254.2022.10.01

Figure Lengend Snippet: SOX2-OT positively modulates SOX2 by sponging miR-200c in H520 cells. A: Bioinformatic analysis of the binding sites of miR-200c on SOX2 and SOX2-OT predicted by StarBase. B: Transcription of SOX2 and SOX2-OT is inhibited by miR-200c mimic and elevated by miR-200c inhibitor. C: miR- 200c inhibitor counteracts SOX2-OT knockdown-induced downregulation of SOX2 and SOX2-OT. D: Expressions of SOX2 and SOX2-OT are positively correlated based on data from cBioPortal database. E: SOX2 overexpression attenuated SOX2-OT knockdown-induced inhibition of GLI1 expression. **P < 0.01.

Article Snippet: Short hairpin RNA (shRNA) plasmids against SOX2-OT were purchased from Genechem (Shanghai, China).

Techniques: Binding Assay, Over Expression, Inhibition, Expressing