Journal: Journal of cellular and molecular medicine

Article Title: Aryl Hydrocarbon Receptor Alleviates Hepatic Fibrosis by Inducing Hepatic Stellate Cell Ferroptosis.

doi: 10.1111/jcmm.70278

Figure Lengend Snippet: FIGURE 1 | AHR directly regulates Mrp1 transcription in mHSCs. (A–C) The expression of Ahr, Mrp1 or Cyp1a1 was detected by QPCR. (D) There are two potential AHR exogenous response elements on the Mrp1 promoter sequence. (E) Promoter sequences containing XREL1 and XREL2 were cloned onto PGL3 plasmids, and mutant plasmids were constructed. (F) Double luciferase assay to detect luciferase activity. (G) EMSA detects AHR binding to specific elements XREL1 and XREL2. (H) CHIP detects AHR binding to specific elements XREL1 and XREL2. Data are expressed as means ± SD; *p < 0.05, **p < 0.01 and ***p < 0.001; Student's t-test or one-way ANOVA.

Article Snippet: ChIP assays were performed using the ChIP Assay kit (Beyotime, P2078). mHSCs were treated with YH439 (5 μM) for 24 h. Subsequently, mHSCs were sonicated and then immunoprecipitated with the antibody against AHR (1:100 dilution BOSTER Cat# A00225- 4, RRID: AB_3095576) with IgG (1:100 dilution Proteintech Cat# 30000- 0- AP RRID AB_2819035) as a negative control.

Techniques: Expressing, Sequencing, Clone Assay, Mutagenesis, Construct, Luciferase, Activity Assay, Binding Assay

Journal: Journal of visualized experiments : JoVE

Article Title: A Mouse Model to Investigate the Role of Cancer-associated Fibroblasts in Tumor Growth

doi: 10.3791/61883

Figure Lengend Snippet: 2. Generate cancer cells for injection

Article Snippet: Aliquot resuspended cells onto new tissue culture plates of the appropriate size with DMEM + 10% FBS (10 mL of media is suitable for a 10 cm tissue culture plate). table ft1 table-wrap mode="anchored" t5 Name of Material/Equipment Company Catalog Number Comments/Description Centrifuge for conical tubes capable of reaching 180 × g Fisher Scientific 14-432-22 Sterle 25 ml serological pipet Celltreat 667225B Sterile 10 ml serological pipet Celltreat 667210B Sterile 5 ml serological pipet Celltreat 229005B Sterile 50 ml centrifuge tubes Genesee Scientific 28-108 TC treated 150 × 20 mm dishes Genesee Scientific 25-203 TC treated 100 × 20 mm dishes Genesee Scientific 25-202 TC treated 60 × 15 mm dishes Genesee Scientific 25-260 Cell Culture Multi Flasks Fisher Scientific 14-826-95 Dulbecco’s Modified Eagle Medium Fisher Scientific 11965-118 Sterile tissue-culture grade PBS Fisher Scientific 50-751-7476 Sterile tissue culture-grade Trypsin-EDTA Fisher Scientific 15400054 Fetal bovine serum Fisher Scientific MT35010CV Countess Cell Counting Chamber Fisher Scientific C10228 Cancer cells ATCC ATCC® CRL-11147™ This is the catalog number for a primary human melanoma cell line.

Techniques: Cell Culture, Modification, Cell Counting, Derivative Assay, Injection