Journal: Cell division

Article Title: Long non-coding RNA NEAT1 induced by BHLHE40 activates Wnt/β-catenin signaling and potentiates colorectal cancer progression.

doi: 10.1186/s13008-024-00129-7

Figure Lengend Snippet: Fig. 2 BHLHE40 mediates the transcription of NEAT1 in CRC cells. A Genes co-expressed with NEAT1 in CRC were obtained from UALCAN. B The intersection of genes co-expressed with NEAT1 in COAD and READ and the transcription factors targeting NEAT1 downloaded from hTFtarget. C The expression of these eight transcription factors in CRC was analyzed in the UALCAN. D The expression of BHLHE40 in both COAD and READ. E The binding fragment of BHLHE40 on the NEAT1 promoter with the highest score. F RT-qPCR detection of BHLHE40 expression in CRC tissues and adjacent tissues by RT-qPCR (n = 54). G Expression of NEAT1 in CRC cell lines and HCoEpiC cells by RT-qPCR. H Knockdown efficiency of BHLHE40 by RT-qPCR. I RT-qPCR detection of NEAT1 expression after knockdown of BHLHE40 in LoVo and HCT-15 cells. J Changes in luciferase activity after knockdown of BHLHE40 using luciferase activity assay. K BHLHE40 binding to the NEAT1 promoter in CRC cell using ChIP assay. Results were expressed as magnitude of relative expression (means ± SD) from three independent experiments. F Paired t-test; G one-way ANOVA; H–K two-way ANOVA. *p < 0.05

Article Snippet: Afterward, the membranes were sealed with non-fat milk at room temperature for 2 h. Primary antibodies to BHLHE40 (1:1000, 17895-1-AP, ProteinTech Group, Chicago, IL, USA), Wnt (1:500, 27935-1-AP, ProteinTech Group), β-catenin (1:2000, 17565-1-AP, ProteinTech), cyclin D1 (1:5000, 26939-1-AP, ProteinTech), c-myc (1:500, GTX103436, GeneTex, Inc., Alton Pkwy Irvine, CA, USA), E-cadherin (1:500, GTX100443, GeneTex), N-cadherin (1:500, GTX127345, GeneTex), Vimentin (1:5000, GTX100619, GeneTex), and β-actin (1:500, D110001, Sangon) were applied overnight at 4 °C.

Techniques: Expressing, Binding Assay, Quantitative RT-PCR, Knockdown, Luciferase, Activity Assay

Journal: Cell division

Article Title: Long non-coding RNA NEAT1 induced by BHLHE40 activates Wnt/β-catenin signaling and potentiates colorectal cancer progression.

doi: 10.1186/s13008-024-00129-7

Figure Lengend Snippet: Fig. 4 NEAT1 reverses the metastasis-suppressing and tumor-suppressing properties of sh-BHLHE40. HCT-15 cells after infection were injected subcutaneously into nude mice to observe tumor growth. A Tumor growth curve. B Immunohistochemical analysis of BHLHE40 and Ki67 expression in xenograft tumor tissues of nude mice. C HCT-15 cells after infection were injected into nude mice via the tail vein, and lung metastasis formation was observed using HE staining. D Western blot for BHLHE40 and EMT-related protein expression in metastatic tissues with lung infiltration. Results were expressed as magnitude of relative expression (means ± SD) from six nude mice in each group. Two-way ANOVA. *p < 0.05

Article Snippet: Afterward, the membranes were sealed with non-fat milk at room temperature for 2 h. Primary antibodies to BHLHE40 (1:1000, 17895-1-AP, ProteinTech Group, Chicago, IL, USA), Wnt (1:500, 27935-1-AP, ProteinTech Group), β-catenin (1:2000, 17565-1-AP, ProteinTech), cyclin D1 (1:5000, 26939-1-AP, ProteinTech), c-myc (1:500, GTX103436, GeneTex, Inc., Alton Pkwy Irvine, CA, USA), E-cadherin (1:500, GTX100443, GeneTex), N-cadherin (1:500, GTX127345, GeneTex), Vimentin (1:5000, GTX100619, GeneTex), and β-actin (1:500, D110001, Sangon) were applied overnight at 4 °C.

Techniques: Infection, Injection, Immunohistochemical staining, Expressing, Staining, Western Blot

Journal: Cell division

Article Title: Long non-coding RNA NEAT1 induced by BHLHE40 activates Wnt/β-catenin signaling and potentiates colorectal cancer progression.

doi: 10.1186/s13008-024-00129-7

Figure Lengend Snippet: Fig. 5 BHLHE40 mediates NEAT1 transcription to activate Wnt/β-catenin signaling in CRC cells. A Western blot for the E-cadherin, N-cadherin, Vimentin, Wnt, β-catenin, c-myc, and cyclin D1 protein expression in CRC cells in response to oe-BHLHE40 (oe-NC as control) or oe-BHLHE40 + sh-NEAT1 (oe-BHLHE40 + sh-NC as control). B Effects of oe-BHLHE40 and iCRT3 combined treatment on Wnt/β-catenin pathway activity in CRC cells was measured using TOP/FOP flash assay. C MTT assay for LoVo and HCT-15 cell viability. D Transwell assay for LoVo and HCT-15 cell migration ability. E Transwell assay for LoVo and HCT-15 cell invasion ability. F Flow cytometry for LoVo and HCT-15 cell apoptosis. Results were expressed as magnitude of relative expression (means ± SD) from three independent experiments. Two-way ANOVA. *p < 0.05

Article Snippet: Afterward, the membranes were sealed with non-fat milk at room temperature for 2 h. Primary antibodies to BHLHE40 (1:1000, 17895-1-AP, ProteinTech Group, Chicago, IL, USA), Wnt (1:500, 27935-1-AP, ProteinTech Group), β-catenin (1:2000, 17565-1-AP, ProteinTech), cyclin D1 (1:5000, 26939-1-AP, ProteinTech), c-myc (1:500, GTX103436, GeneTex, Inc., Alton Pkwy Irvine, CA, USA), E-cadherin (1:500, GTX100443, GeneTex), N-cadherin (1:500, GTX127345, GeneTex), Vimentin (1:5000, GTX100619, GeneTex), and β-actin (1:500, D110001, Sangon) were applied overnight at 4 °C.

Techniques: Western Blot, Expressing, Control, Activity Assay, MTT Assay, Transwell Assay, Migration, Flow Cytometry

Journal: iScience

Article Title: Basic-helix-loop-helix family member e41 suppresses osteoclastogenesis and abnormal bone resorption disease via NFATc1.

doi: 10.1016/j.isci.2024.109059

Figure Lengend Snippet: Figure 3. BHLHE41 regulates osteoclast differentiation via NFATc1 (A) Heatmap of differentially expressed genes (DEGs) for BMMs in si-NC and si-BHLHE41 groups (n = 4). (B and C)Volcano plot and quantification of RNA-seq data of BMMs in si-NC and si-BHLHE41 groups. (D) GOBubble plot of KEGG enrichment analysis in DEGs. (E) GOBubble plot of GO enrichment analysis in DEGs. (F) GOCircle plot of GO enrichment analysis of the DEGs.

Article Snippet: The primary antibodies included: BHLHE41 (Affinity, Cat#AF0442, 1:1000) , NFATc1 (Affinity, Cat#DF6446, 1:1000) or GAPDH (Proteintech Group In, Cat# 60004-1-Ig, 1:4000).

Techniques: RNA Sequencing