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

Journal: Nature Communications

Article Title: Actin dysregulation induces neuroendocrine plasticity and immune evasion: a vulnerability of small cell lung cancer

doi: 10.1038/s41467-025-67078-9

Figure Lengend Snippet: a UMAPs of SCLC tumor cells from 19 SCLC patient tumor samples (54,633 cells) and 8 normal lung samples (24,041 cells). Each dot represents a single cell, colored by a human sample ID (left) and SCLC vs. normal (right). b Correlation matrix plot showing pair-wise correlations among the human normal lung and 19 patient tumor samples. The dendrogram shows the distance of each dataset based on principal component analysis, and Pearson’s correlation is displayed with a color spectrum. Groups of patients were unbiasedly categorized by dendrogram and correlation. c Sankey plot shows the correlation between SCLC subtypes (MS1 and MS2) and clinical information (cancer type and stage). d Dot plot showing NE marker gene expression in 19 SCLC patient samples. ANPY ( A SCL1 , N EUROD1 , P OU2F3 , and Y AP1 )-based classification was noted at the top. e , f Violin ( e ) and dot ( f ) plots showing CRACD mRNA and EZH2/NOTCH-target scores. P values: two-sided Student’s t -test. g Violin plots of MHC-I pathway gene expression in MS1 vs MS2 tumors; two-sided Student’s t -test. h GSEA of gene sets associated with EZH2 targets and the MHC-I pathway in MS1 compared to MS2. i Correlation scatter plots for Pearson’s correlation analysis (using GraphPad Prism) of CRACD and MHC-I genes ( HLA-A , B , C , E , and TAP1/2 ) in SCLC patient tumor cells based on the TCGA bulk RNA-seq datasets. r, Pearson correlation coefficient; P values were calculated using two-sided Student’s t -test. j Illustration of the impact of CRACD loss on SCLC tumorigenesis. CRACD-positive tumors maintain MHC-I–mediated immune activity (“hot tumors”), whereas CRACD-deficient tumors acquire NE plasticity and immune evasion. Loss of CRACD disrupts actin polymerization, activating HIPPO signaling and LATS1/2-mediated YAP1 inhibition, which suppresses NOTCH1 and induces ASCL1-driven NE gene expression. Concurrently, EZH2-dependent silencing of MHC-I genes converts tumors to immune “cold” states, accelerating tumor progression. j was partly created with BioRender.com. Source data are provided as a Source Data file.

Article Snippet: Poly(A)-enriched mRNA libraries were prepared using Novogene’s standard workflow for eukaryotic mRNA (WOBI) and sequenced on an Illumina NovaSeq X Plus platform to generate paired-end 150 bp reads, yielding ~6 Gb of raw data per sample.

Techniques: Marker, Gene Expression, RNA Sequencing, Activity Assay, Inhibition