luciferase  (Addgene inc)

Journal: International Journal of Molecular Medicine

Article Title: miRNA-378a-5p attenuates the development of abdominal aortic aneurysm via ABLIM1-MKL1 signaling pathways

doi: 10.3892/ijmm.2026.5768

Figure Lengend Snippet: ABLIM1 is a downstream target of miR-378a-5p and involves in abdominal aortic aneurysm development. (A) Volcano plot of differently expressed genes in GSE183464 and GSE237229 database. Venn diagram showed intersection of differentially expressed genes and predicted target genes including ABLIM1 , DDX5 and SLC7A1 . (B) RT-qPCR analysis of target genes Ablim1 , Ddx5 and Slc7a1 in the TNFα-treated vascular smooth muscle cells (n=3 per group). RT-qPCR analysis of Ablim1 , Ddx5 and Slc7a1 in the aortas of Ang II-treated mice (n=3 per group). (C) ABLIM1 expression in the aortas treated-with antagomir-NC or antagomir-378a-5p was identified using by immunofluorescence staining. ABLIM1 (red), α-SMA (green) and DAPI (blue). (D) Representative images of immunofluorescence staining identified ABLIM1 expression in aortas with angomir-NC or angomir-378a-5p. ABLIM1 (red), α-SMA (green) and DAPI (blue). (E) Conservatism analysis of the binding site for miR-378a-5p and ABLIM1 in in humans, mice and rat. (F) Luciferase activity in 293T cells transfected with mimics-miR-378a-5p together with ABLIM1 -3'UTR-wild type or mutant plasmid. Data are presented as the mean ± SEM. P-values were calculated by Student's t test (for B). ** P<0.01 vs. Control or saline or mimics-NC- ABLIM1 -3'UTR-WT. ABLIM1, actin-binding LIM protein 1; miR or miRNA, microRNA; RT-qPCR, reverse transcription-quantitative PCR; α-SMA, α-smooth muscle actin; NC, negative control; UTR, untranslated region; WT, wild-type; MUT, mutated.

Article Snippet: The 3' untranslated region (3'UTR) of mouse Ablim1 , Ddx5 (Dead-box helicase 5), Slc7a1 (Cationic amino acid transporter 1) gene was amplified and cloned into the pMIR-REPORT Luciferase (OBiO Technology Company) to construct pMIR-REPORT Luciferase 3'UTR wild-type (WT) plasmids.

Techniques: Quantitative RT-PCR, Expressing, Immunofluorescence, Staining, Binding Assay, Luciferase, Activity Assay, Transfection, Mutagenesis, Plasmid Preparation, Control, Saline, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control

Journal: Immunity, Inflammation and Disease

Article Title: WTAP Contributes to Periodontitis Pathogenesis by Promoting PDLSC Senescence and Impairing Osteogenic Differentiation via m6A‐Dependent Regulation of TP53BP1

doi: 10.1002/iid3.70335

Figure Lengend Snippet: WTAP modifies TP53BP1 via m6A. (a) The m6A sites of TP53BP1 were predicted by the SRAMP website. (b, c) TP53BP1 expression in P‐PDLSCs transfected with shNC and shWTAP was measured by RT‐qPCR and western blot, respectively (Unpaired t ‐test). (d) MeRIP was used to determine the m6A levels on TP53BP1 mRNA in P‐PDLSCs transfected with shNC and shWTAP (Unpaired t ‐test). (e) TP53BP1 mRNA enrichment in P‐PDLSCs treated with anti‐IgG + shNC, anti‐IgG + shWTAP, anti‐WTAP+shNC, and anti‐WTAP + shWTAP was examined by RIP (one‐way ANOVA). (f) TP53BP1 mRNA expression in P‐PDLSCs co‐treated with shNC or shWTAP and Act D for 0, 2, 4, and 6 h was detected by RT‐qPCR (one‐way ANOVA). (g) Luciferase activities in the WT‐TP53BP1 + shNC, WT‐TP53BP1 + shWTAP, MUT‐TP53BP1 + shNC, and MUT‐TP53BP1+shWTAP groups were measured by dual‐luciferase reporter assay (one‐way ANOVA). All experiments were conducted at least three independent times. * p < 0.05.

Article Snippet: Wild‐type (WT) and mutant (MUT) TP53BP1 3′UTR luciferase reporter plasmids (WT‐TP53BP1 and MUT‐TP53BP1) were constructed by Sangon Biotech (Shanghai) Co. Ltd. (Shanghai, China).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot, Luciferase, Reporter Assay

Journal: Immunity, Inflammation and Disease

Article Title: WTAP Contributes to Periodontitis Pathogenesis by Promoting PDLSC Senescence and Impairing Osteogenic Differentiation via m6A‐Dependent Regulation of TP53BP1

doi: 10.1002/iid3.70335

Figure Lengend Snippet: TP53BP1 downregulation mitigates WTAP‐caused P‐PDLSC senescence and oxidative stress. (a) RT‐qPCR was used to measure the mRNA levels of TP53BP1 in P‐PDLSCs transfected with shNC and shTP53BP1 (Unpaired t ‐test). (b) Western blot was applied to determine the protein levels of TP53BP1 in P‐PDLSCs transfected with shNC and shTP53BP1 (Unpaired t ‐test). (c–h) The P‐PDLSCs were treated with oeNC + shNC, oeWTAP + shNC, and oeWTAP + shTP53BP1. (c–e) The MDA, SOD, and ROS levels were examined using the corresponding kits (one‐way ANOVA). (f) The γ‐H2AX positive cells were detected using IF (Scale bar = 100 μm; one‐way ANOVA). (g) SA‐β‐gal positive cells were measured using the corresponding kit (Scale bar = 100 μM; one‐way ANOVA). (h) Western blot was applied to determine the protein levels of p53 and p16 (one‐way ANOVA). All experiments were conducted at least three independent times. * p < 0.05.

Article Snippet: Wild‐type (WT) and mutant (MUT) TP53BP1 3′UTR luciferase reporter plasmids (WT‐TP53BP1 and MUT‐TP53BP1) were constructed by Sangon Biotech (Shanghai) Co. Ltd. (Shanghai, China).

Techniques: Quantitative RT-PCR, Transfection, Western Blot

Journal: Immunity, Inflammation and Disease

Article Title: WTAP Contributes to Periodontitis Pathogenesis by Promoting PDLSC Senescence and Impairing Osteogenic Differentiation via m6A‐Dependent Regulation of TP53BP1

doi: 10.1002/iid3.70335

Figure Lengend Snippet: WTAP represses the osteogenic differentiation of P‐PDLSCs via TP53BP1. The P‐PDLSCs were treated with oeNC+shNC, oeWTAP+shNC, and oeWTAP + shTP53BP1. (a) ALP staining of P‐PDLSCs induced for 7 days (Scale bar = 100 μM; one‐way ANOVA). (b) ARS staining of P‐PDLSCs induced for 28 days (Scale bar = 100 μM; one‐way ANOVA). (c) Western blot was used to determine the protein levels of OPN, OCN, and RUNX2 (one‐way ANOVA). All experiments were conducted at least three independent times. * p < 0.05.

Article Snippet: Wild‐type (WT) and mutant (MUT) TP53BP1 3′UTR luciferase reporter plasmids (WT‐TP53BP1 and MUT‐TP53BP1) were constructed by Sangon Biotech (Shanghai) Co. Ltd. (Shanghai, China).

Techniques: Staining, Western Blot

Journal: bioRxiv

Article Title: Systematic identification and characterization of regulators of aryl hydrocarbon receptor signaling

doi: 10.1101/2025.09.15.676391

Figure Lengend Snippet: (A) log 2 fold change in luciferase signal after treatment with the indicated chemicals for 4 h relative to vehicle control. (B) Renilla relative luminescence units (RLUs, average of duplicate treatments) normalized to mean Renilla RLUs across the entire library plotted against firefly RLUs (average of duplicate treatments) for all chemicals in the 250-chemical library. All chemicals with fraction mean Renilla RLUs ≤ 0.5 are excluded from analysis. (C) Dose-response curves of the two additional urolithins that activate AHR, after treatment of the HepG2 luciferase reporter cells for 4 h. (D) Dose-response curves of 3 additional chemicals from the 250-chemical screen that induce AHR activation, after treatment of the HepG2 luciferase reporter cells for 4 h. Data represent treatment replicates ( n =2).

Article Snippet: The second expression cassette contains a UBC constitutive promoter followed by a renilla luciferase (cloned from Addgene #113088), P2A, and eGFP.

Techniques: Luciferase, Control, Activation Assay