Journal: Oncogenesis

Article Title: Investigation of lncRNA expression in newly diagnosed multiple myeloma reveals a LINC01432-CELF2 axis as an inhibitor of apoptosis

doi: 10.1038/s41389-025-00579-w

Figure Lengend Snippet: a POSTAR3-predicted CELF2 binding sites on LINC01432 lncRNA. b Comprehensive identification of RNA-binding proteins by mass spectrometry (ChIRP-ms) followed by western blot analysis demonstrating enrichment of CELF2 protein in LINC01432 pulldown fraction, as compared to unprocessed input lysate, with GAPDH negative control in RPMI 8226 and OPM2 cells. c Schematic of CELF2-binding sites on LINC01432 with locations of site-specific primers used for iCLIP RT-qPCR. Created with Biorender.com. CELF2 iCLIP RT-qPCR showing enrichment of LINC01432 binding in d RPMI 8826 and e OPM2 cells, as compared to IgG and GAPDH negative controls. RNA immunoprecipitation (RIP) followed by RT-qPCR in nuclear and cytoplasmic compartments indicating LINC01432 enrichment in CELF2 immunoprecipitants in f RPMI 8826 and g OPM2 cells. h Multiplex fluorescent in situ hybridization (mFISH) for LINC01432 combined with CELF2 immunohistochemistry in bone marrow aspirates from newly diagnosed multiple myeloma (NDMM) patients. i Quantification of subcellular localization of LINC01432 and CELF2 in NDMM samples using QuPath analysis. Scale bar = 40 µM *fold enrichment >1.5, **fold enrichment >5, ***fold enrichment >10.

Article Snippet: Interactions with LINC01432 were assessed using CELF2 antibody and anti-GAPDH antibody (Cell Signaling, Danvers, MA, USA, catalog number 97166S).

Techniques: Binding Assay, RNA Binding Assay, Mass Spectrometry, Western Blot, Negative Control, Quantitative RT-PCR, RNA Immunoprecipitation, Multiplex Assay, In Situ Hybridization, Immunohistochemistry

Journal: Oncogenesis

Article Title: Investigation of lncRNA expression in newly diagnosed multiple myeloma reveals a LINC01432-CELF2 axis as an inhibitor of apoptosis

doi: 10.1038/s41389-025-00579-w

Figure Lengend Snippet: a Heatmap of RNA sequencing data illustrating expression profiles of overlapping genes across Control LNA ASOs, LINC01432 -targeted LNA ASOs, and CELF2-targeted LNA ASOs. b Venn diagram depicting significantly overlapping differentially expressed genes (adjusted p < 0.05, |log₂FC|> 1.5). c Distribution of gene biotypes among the shared targets, including long non-coding RNAs (lncRNAs), protein-coding genes, and other RNA classes. d Top overlapping significantly dysregulated genes following CELF2 and LINC01432 knockdown. e Overlapping lncRNAs with most highly altered expression following CELF2 and LINC01432 knockdown. f Shared genes with established roles in multiple myeloma pathogenesis. *log fold change >2, **log fold change >3, ***log fold change >4, #log fold change >5.

Article Snippet: Interactions with LINC01432 were assessed using CELF2 antibody and anti-GAPDH antibody (Cell Signaling, Danvers, MA, USA, catalog number 97166S).

Techniques: RNA Sequencing, Expressing, Control, Knockdown

Journal: Oncogenesis

Article Title: Investigation of lncRNA expression in newly diagnosed multiple myeloma reveals a LINC01432-CELF2 axis as an inhibitor of apoptosis

doi: 10.1038/s41389-025-00579-w

Figure Lengend Snippet: We identified 248 deregulated long non-coding RNAs (lncRNAs) in newly diagnosed multiple myeloma patient samples with short progression-free survival (PFS) in response to standard MM therapy, as compared to those with prolonged PFS. Among these, LINC01432 was the most significantly upregulated and was found to interact with the RNA-binding protein CELF2. Elevated expression of LINC01432 , or activation of the LINC01432 –CELF2 regulatory axis, modulates gene networks involved in chemokine and cytokine signaling as well as interferon-stimulated pathways, collectively promoting cell survival and inhibiting apoptosis. Created with Biorender.com.

Article Snippet: Interactions with LINC01432 were assessed using CELF2 antibody and anti-GAPDH antibody (Cell Signaling, Danvers, MA, USA, catalog number 97166S).

Techniques: RNA Binding Assay, Expressing, Activation Assay

Journal: Scientific Reports

Article Title: The regulation of miR-155 strand selection by CELF2, FUBP1 and KSRP proteins

doi: 10.1038/s41598-025-15004-w

Figure Lengend Snippet: CELF2, FUBP1, and KSRP deficiency alters the expression of miR-155-5p and miR-155-3p in response to LPS ± IL10 stimulation. FUBP1 KD, CELF2 KD, KSRP KD or the control RAW264.7 cells were stimulated with 1 ng/mL LPS ± 1 ng/mL of IL10 for ( A ) 2 and 4 h before total RNA extraction. The expression levels of miR-155-5p and miR-155-3p were determined by qPCR and normalized to snoRNA202 levels. The data plotted represents the expression levels of miR-155-5p and miR-155-3p levels normalized to the 2-hour LPS-stimulated control RAW264.7 cells. ( B ) The data from panel A were replotted as time courses for each cell type. miR-155-5p and miR-155-3p levels were normalized to the unstimulated sample of control RAW264.7 cells. Two-way ANOVA with Tukey’s correction determined the comparisons indicated with braces between different time points or cell types as * and the significance between stimulations as †. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05, †††† p < 0.0001, ††† p < 0.001, †† p < 0.01, † p < 0.05, ns = not significant. The data are representative of 4 experiments.

Article Snippet: The membranes were blocked in 3% bovine serum albumin (BSA), then probed with the following primary antibodies overnight: 1:1000 KSRP (ab140648, Abcam, Toronto, ON), 0.1 μg/mL GAPDH (G9545, Millipore-Sigma, Oakville, ON), 1 μg/mL CELF2 (Sc-47731, Santa Cruz, Dallas, TX) and 1 μg/mL FUBP1 or KSRP (Sc-136137, Santa Cruz, Dallas, TX).

Techniques: Expressing, Control, RNA Extraction

Journal: Scientific Reports

Article Title: The regulation of miR-155 strand selection by CELF2, FUBP1 and KSRP proteins

doi: 10.1038/s41598-025-15004-w

Figure Lengend Snippet: Expression of miR-155 target genes in CELF2 KD, FUBP1 KD, KSRP KD, or control RAW264.7 cells. Cells were stimulated with 1 ng/mL of LPS for 4 hours before extraction of total cellular RNA. The expression levels of Arg2, Apbb2 and Myd88 were determined by qPCR and normalized to GAPDH levels. Two-way ANOVA with Tukey’s correction determined the comparisons indicated with braces. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05, ns = not significant. The data represents three experiments.

Article Snippet: The membranes were blocked in 3% bovine serum albumin (BSA), then probed with the following primary antibodies overnight: 1:1000 KSRP (ab140648, Abcam, Toronto, ON), 0.1 μg/mL GAPDH (G9545, Millipore-Sigma, Oakville, ON), 1 μg/mL CELF2 (Sc-47731, Santa Cruz, Dallas, TX) and 1 μg/mL FUBP1 or KSRP (Sc-136137, Santa Cruz, Dallas, TX).

Techniques: Expressing, Control, Extraction

Journal: Scientific Reports

Article Title: The regulation of miR-155 strand selection by CELF2, FUBP1 and KSRP proteins

doi: 10.1038/s41598-025-15004-w

Figure Lengend Snippet: Schematic representation of FUBP1/KSRP/CELF2 regulation, homology, and interaction. ( A ) The sequence alignment of the KH3 domain of FUBP1 and KSRP protein. Sequences in black and red indicate matching and mismatching sequences, respectively. ( B ) Schematic diagram showing the predicted interaction site of FUBP1, CELF2 and KSRP to pre-miR-155. The bases of pre-miR-155 are gray; blue bases show the miR-155-5p sequence, and black bases indicate the miR-155-3p sequence.

Article Snippet: The membranes were blocked in 3% bovine serum albumin (BSA), then probed with the following primary antibodies overnight: 1:1000 KSRP (ab140648, Abcam, Toronto, ON), 0.1 μg/mL GAPDH (G9545, Millipore-Sigma, Oakville, ON), 1 μg/mL CELF2 (Sc-47731, Santa Cruz, Dallas, TX) and 1 μg/mL FUBP1 or KSRP (Sc-136137, Santa Cruz, Dallas, TX).

Techniques: Sequencing

Journal: Scientific Reports

Article Title: The regulation of miR-155 strand selection by CELF2, FUBP1 and KSRP proteins

doi: 10.1038/s41598-025-15004-w

Figure Lengend Snippet: Increased interaction of FUBP1 protein with pre-miR-155 in response to LPS stimulation. RAW264.7 cells were transfected with biotinylated pre-miR-155 oligonucleotide and stimulated with LPS ± IL10 for 1 h before collecting the samples. ( A ) Expression levels of CELF2, FUBP1 and KSRP protein interacting with pre-miR-155 oligonucleotide were determined by immunoblotting. ( B )The graphs show the CELF2, FUBP1, and KSRP band intensities in the pull-down sample, normalized to the same protein in the total cell lysate. One-way ANOVA with Tukey’s correction calculated the significance of the difference between the stimulations. ** p < 0.01, * p < 0.05, ns = not significant. The data represent the results of 5 experiments.

Article Snippet: The membranes were blocked in 3% bovine serum albumin (BSA), then probed with the following primary antibodies overnight: 1:1000 KSRP (ab140648, Abcam, Toronto, ON), 0.1 μg/mL GAPDH (G9545, Millipore-Sigma, Oakville, ON), 1 μg/mL CELF2 (Sc-47731, Santa Cruz, Dallas, TX) and 1 μg/mL FUBP1 or KSRP (Sc-136137, Santa Cruz, Dallas, TX).

Techniques: Transfection, Expressing, Western Blot