Journal: Animals : an Open Access Journal from MDPI

Article Title: Pleiotropic Gene HMGA2 Regulates Myoblast Proliferation and Affects Body Size of Sheep

doi: 10.3390/ani14182721

Figure Lengend Snippet: Identification of HMGA2 core promoter and variation. ( A ) Relative luciferase activities of different lengths of promoter fragments; ( B ) Sanger sequencing detected SNP rs428001129 in NC_056056.1: 154134300-154134882 (-618 to -1200 bp upstream of the HMGA2 TSS) containing a core promoter; ( C ) rs428001129 changed HMGA2 promoter activity. P1, NC_056056.1: 154133741-154135682 (upstream of TSS, -59 to -2000 bp); P2, NC_056056.1: 154133741-154135281 (upstream of TSS, -59 to -1599 bp); P3, NC_056056.1: 154133741-154134882 (upstream of TSS, -59 to -1200 bp); P4, NC_056056.1: 154133741-154134300 (upstream of TSS, -59 to -618 bp). ** p < 0.01.

Article Snippet: In this study, a series of promoter deletion fragments of HMGA2 were cloned into the promoter region of the firefly luciferase gene of the pGL3-Basic vector (Addgene #212936) with the Renilla luciferase of the pRL-TK vector as an internal control.

Techniques: Luciferase, Sequencing, Activity Assay

Journal: Cell Death & Disease

Article Title: Downregulated liver-elevated long intergenic noncoding RNA (LINC02428) is a tumor suppressor that blocks KDM5B/IGF2BP1 positive feedback loop in hepatocellular carcinoma

doi: 10.1038/s41419-023-05831-y

Figure Lengend Snippet: A FISH assay showed that LINC02428 was mainly located in the cytoplasm of HCC cells . B Western blotting of proteins obtained by RNA pull-down assay showed that LINC02428 bound to IGF2BP1. C RIP assays confirmed that LINC02428 interacts with IGF2BP1. D FISH and IF assays show LINC02428 and IGF2BP1 are co-localized in the cytoplasm in HCC cells. E Deletion mapping of the IGF2BP1-binding domain in LINC02428 was examined by western blotting. F qRT-PCR showed LINC02428 upregulation decreased IGF2BP1 mRNA levels. G Western blotting indicated LINC02428 overexpression decreased IGF2BP1 protein levels.

Article Snippet: Firefly luciferase reporter plasmid of IGF2BP1(pGL3-Basic-IGF2BP1-Pro-5′UTR) and ranilla luciferase reporter plasmid (pRL-TK) were co-transfected with overexpression plasmid of KDM5B or control vector to examine luciferase activities by Dual-Luciferase Reporter Gene Assay Kit (Genepharma, China).

Techniques: Western Blot, Pull Down Assay, Binding Assay, Quantitative RT-PCR, Over Expression

Journal: Cell Death & Disease

Article Title: Downregulated liver-elevated long intergenic noncoding RNA (LINC02428) is a tumor suppressor that blocks KDM5B/IGF2BP1 positive feedback loop in hepatocellular carcinoma

doi: 10.1038/s41419-023-05831-y

Figure Lengend Snippet: A Increased LINC02428 expression resulted in decreased KDM5B mRNA expression. B KDM5B downregulation decreased IGF2BP1 mRNA levels. C The protein lysate of KDM5B mRNA pull-down contained IGF2BP1. D RIP assays showed that IGF2BP1 could recruit KDM5B mRNA. E RNA stability assay indicated that LINC02428 overexpression could weaken the stability of KDM5B mRNA. F IGF2BP1 upregulation could reverse the LINC02428-mediated decrease in KDM5B mRNA stability. G ChIP-qPCR assay confirmed that KDM5B enriched in the promoter of IGF2BP1 and LINC02428 upregulation could decrease KDM5B enrichment. H Dual luciferase reporter assay proved that KDM5B bound to IGF2BP1 promoter to promote its transcription, and IGF2BP1 transcription promoted KDM5B could be enhanced by upregulation of IGF2BP1 or decrease by LINC02428 overexpression in 293 T. I Western blotting showed that KDM5B silencing could decrease expression of IGF2BP1 protein in HCC cells.

Article Snippet: Firefly luciferase reporter plasmid of IGF2BP1(pGL3-Basic-IGF2BP1-Pro-5′UTR) and ranilla luciferase reporter plasmid (pRL-TK) were co-transfected with overexpression plasmid of KDM5B or control vector to examine luciferase activities by Dual-Luciferase Reporter Gene Assay Kit (Genepharma, China).

Techniques: Expressing, Stability Assay, Over Expression, Luciferase, Reporter Assay, Western Blot

Journal: Cell Death & Disease

Article Title: Downregulated liver-elevated long intergenic noncoding RNA (LINC02428) is a tumor suppressor that blocks KDM5B/IGF2BP1 positive feedback loop in hepatocellular carcinoma

doi: 10.1038/s41419-023-05831-y

Figure Lengend Snippet: A–C EdU assay D CCK-8 assay indicated that KDM5B silencing could suppress HCC proliferation; however, IGF2BP1 upregulation could reverse the inhibition of proliferation. E–G IGF2BP1 overexpression rescued the KDM5B silencing-induced inhibition of migration and invasion in HCC cells. H Western blotting confirmed KDM5B knockdown downregulated IGF2BP1, whereas IGF2BP1 co-transfection reversed KDM5B downregulation.

Article Snippet: Firefly luciferase reporter plasmid of IGF2BP1(pGL3-Basic-IGF2BP1-Pro-5′UTR) and ranilla luciferase reporter plasmid (pRL-TK) were co-transfected with overexpression plasmid of KDM5B or control vector to examine luciferase activities by Dual-Luciferase Reporter Gene Assay Kit (Genepharma, China).

Techniques: EdU Assay, CCK-8 Assay, Inhibition, Over Expression, Migration, Western Blot, Cotransfection

Journal: Cell Death & Disease

Article Title: Downregulated liver-elevated long intergenic noncoding RNA (LINC02428) is a tumor suppressor that blocks KDM5B/IGF2BP1 positive feedback loop in hepatocellular carcinoma

doi: 10.1038/s41419-023-05831-y

Figure Lengend Snippet: EdU and CCK-8 assays showed that the upregulation of IGF2BP1 ( A , C , D ) or KDM5B ( B , E , F ) impaired the overexpression of the LINC02428-induced suppression of HCC proliferation. Migration and invasion assays indicated that IGF2BP1 ( G ) or KDM5B ( H ) overexpression rescued the inhibition of HCC migration and invasion induced by LINC02428 upregulation. I Western blotting showed expression of protein about both upregulation LINC02428 and IGF2BP1. J Western blot assays showed protein levels after LINC02428 and KDM5B upregulation.

Article Snippet: Firefly luciferase reporter plasmid of IGF2BP1(pGL3-Basic-IGF2BP1-Pro-5′UTR) and ranilla luciferase reporter plasmid (pRL-TK) were co-transfected with overexpression plasmid of KDM5B or control vector to examine luciferase activities by Dual-Luciferase Reporter Gene Assay Kit (Genepharma, China).

Techniques: CCK-8 Assay, Over Expression, Migration, Inhibition, Western Blot, Expressing

Journal: Cell Death & Disease

Article Title: Downregulated liver-elevated long intergenic noncoding RNA (LINC02428) is a tumor suppressor that blocks KDM5B/IGF2BP1 positive feedback loop in hepatocellular carcinoma

doi: 10.1038/s41419-023-05831-y

Figure Lengend Snippet: LINC02428 blocked the KDM5B/IGF2BP1 positive feedback loop to inhibit HCC progression.

Article Snippet: Firefly luciferase reporter plasmid of IGF2BP1(pGL3-Basic-IGF2BP1-Pro-5′UTR) and ranilla luciferase reporter plasmid (pRL-TK) were co-transfected with overexpression plasmid of KDM5B or control vector to examine luciferase activities by Dual-Luciferase Reporter Gene Assay Kit (Genepharma, China).

Techniques: