Journal: Cancers

Article Title: L1CAM Promotes Human Endometrial Cancer Via NF-κB Activation

doi: 10.3390/cancers18020198

Figure Lengend Snippet: L1CAM is expressed at various levels in human endometrial cancer cells. ( A ) mRNA levels of L1CAM in Ishikawa, HEC-1, HHUA, SPAC-1-L, SPAC-1-S, HOUA. GAPDH was used as an internal control. ( B ) Western blotting of L1CAM and GAPDH in Ishikawa, HEC-1, HHUA, SPAC-1-L, SPAC-1-S, and HOUA. GAPDH was used as a loading control. ( C ) mRNA levels of L1CAM in HHUA and SPAC-1-L with control or L1CAM knockdown. GAPDH was used as an internal control. ( D ) Western blotting of L1CAM and GAPDH in HHUA and SPAC-1-L with control or L1CAM knockdown. GAPDH was used as a loading control. ( E ) Pictures of HHUA and SPAC-1-L with control or L1CAM knockdown. Red arrowheadshows morphological changes ( p < 0.01 **, p < 0.001 ***). The uncropped blots are shown in .

Article Snippet: Poly(A)-based mRNA enrichment was performed using the NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat No. E7490, New England Biolabs, Ipswich, MA, USA).

Techniques: Control, Western Blot, Knockdown

Journal: Cancers

Article Title: L1CAM Promotes Human Endometrial Cancer Via NF-κB Activation

doi: 10.3390/cancers18020198

Figure Lengend Snippet: L1CAM overexpression induces the opposite effects of knockdown. ( A ) mRNA levels of L1CAM in HHUA, SPAC-1-L, and Ishikawa with control or L1CAM overexpression. GAPDH was used as an internal control. ( B ) Western blotting of FLAG, L1CAM, and GAPDH in HHUA, SPAC-1-L, and Ishikawa with control or L1CAM overexpression. GAPDH was used as a loading control. ( C ) MTT assay of HHUA and Ishikawa with control or L1CAM overexpression. ( D ) Cell cycle assay of HHUA and Ishikawa with control or L1CAM overexpression ( p < 0.05 *, p < 0.01 **, p < 0.001 ***, n.s.: not significant). The uncropped blots are shown in .

Article Snippet: Poly(A)-based mRNA enrichment was performed using the NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat No. E7490, New England Biolabs, Ipswich, MA, USA).

Techniques: Over Expression, Knockdown, Control, Western Blot, MTT Assay, Cell Cycle Assay

Journal: Cancers

Article Title: L1CAM Promotes Human Endometrial Cancer Via NF-κB Activation

doi: 10.3390/cancers18020198

Figure Lengend Snippet: L1CAM knockdown suppresses the NF-κB signaling pathway. ( A ) Western blotting of L1CAM, pNF-κB (p65), NF-κB (p65), p-ERK1/2, ERK and GAPDH in HHUA and SPAC-1-L with control or L1CAM knockdown. GAPDH was used as a loading control. ( B ) Relative luciferase activity driven by the NF-κB promoter in HHUA cells with control or L1CAM knockdown. ( C ) Relative mRNA levels of TNF and LTB in HHUA and SPAC-1-L with control or L1CAM knockdown. GAPDH was used as an internal control. ( p < 0.05 *, p < 0.01 **, p < 0.001 ***). The uncropped blots are shown in .

Article Snippet: Poly(A)-based mRNA enrichment was performed using the NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat No. E7490, New England Biolabs, Ipswich, MA, USA).

Techniques: Knockdown, Western Blot, Control, Luciferase, Activity Assay

Journal: Cancers

Article Title: L1CAM Promotes Human Endometrial Cancer Via NF-κB Activation

doi: 10.3390/cancers18020198

Figure Lengend Snippet: L1CAM overexpression activates the NF-kB signaling pathway. ( A ) Relative luciferase activities driven by the NF-κB and TNF promoters in Ishikawa cells. ( B ) Relative luciferase activities driven by the NF-κB promoter and relative TNF mRNA levels in HHUA cells. GAPDH was used as an internal control. ( C ) Western blotting of FLAG, L1CAM, pNF-κB (p65), NF-κB (p65) and GAPDH in Ishikawa and HHUA with control or L1CAM overexpression.GAPDH was used as a loading control. ( p < 0.05 *, p < 0.01 **, p < 0.001 ***). The uncropped blots are shown in .

Article Snippet: Poly(A)-based mRNA enrichment was performed using the NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat No. E7490, New England Biolabs, Ipswich, MA, USA).

Techniques: Over Expression, Luciferase, Control, Western Blot

Journal: Genome Biology and Evolution

Article Title: Landscape of Fluid Sets of Hairpin-Derived 21-/24-nt-Long Small RNAs at Seed Set Uncovers Special Epigenetic Features in Picea glauca

doi: 10.1093/gbe/evw283

Figure Lengend Snippet: Comparisons of DCL3 homologs (A and B), pgRTL2 relative expression (C) and RPM scaled by the number of unique reads (D). Note : gene trees for DCL3 homologs in gymnosperms and several model angiosperms (A); components of DCL3 domains in Arabidopsis (general), Phoenix canariensis and Picea glauca (B); PAZ and dsRB represent Piwi–Argonaute–Zwille and double stranded RNA-binding domains, respectively; thick dashed line in (B) means the upstream of incomplete pgRTL2 mRNA may contain PAZ domain after mapped to its genome (PG29-v.4); RPM represents reads per million and if the absolute expression of the 24-nt sRNA exceeds 2,000 copies in a single library, it is not used for the calculation of RPM and RPM per unique read number. We excluded around 4-5 sRNA reads per library (see supplementary fig. S4 C , Supplementary Material online).

Article Snippet: To enrich sRNAs, total RNA samples underwent polyA selection using Miltenyi MultiMACS mRNA isolation kit (cat. 130-092-519) following the manufacturer’s protocol and the flowthrough (i.e., containing sRNA species without mRNA) was used for plate-based sRNA construction.

Techniques: Expressing, RNA Binding Assay

Journal: Scientific Reports

Article Title: Development and characterization of an inducible Tensin1 deficient transgenic murine model

doi: 10.1038/s41598-026-41319-3

Figure Lengend Snippet: Generation of Tns1 lox, lox mouse for conditional TNS1 knockout. ( A ) Schematic of Tns1 gene locus on Chromosome 1. The targeted exon is expressed in five commonly translated variants. ( B ) Schematic of targeted exon with approximate location of LoxP sites, surrounding exons, and polymerase chain reaction (PCR) primers. C-H. Rosa Cre/+ ; Tns1 lox/lox (Tns1-/-) or TNS1-expressing wild type controls (Tns1+/+) from either B6.Tns1 lox, lox or Rosa Cre/+ ; Tns1 +/+ were treated with daily tamoxifen. ( C ) Gel electrophoresis of tail DNA PCR product confirms excised exon. Real time quantitative PCR of Tns1 ( D ), Tns2 ( E ), and Tns3 ( F ) mRNA expression in lung tissue. Western blot ( G ) and associated densitometry ( H ) in lung tissue ( n = 3 biological repeats). Unpaired Student’s t -test was utilized for all statistical assessments (* p < 0.05). Data is presented as mean ± SD.

Article Snippet: Bulk RNA sequencing analysis was then performed using the Illumina NovaSeq6000 S4 System (2 × 150 bp, ~ 66 M reads/sample) with the Illumina TruSeq Stranded mRNA (polyA enrichment) Library.

Techniques: Knock-Out, Polymerase Chain Reaction, Expressing, Nucleic Acid Electrophoresis, Real-time Polymerase Chain Reaction, Western Blot