Journal: Molecular immunology

Article Title: Chemerin-Activated Functions of CMKLR1 are Regulated by G Protein-Coupled Receptor Kinase 6 (GRK6) and β-Arrestin 2 in Inflammatory Macrophages

doi: 10.1016/j.molimm.2018.12.016

Figure Lengend Snippet: β-arrestin-2 deficient (Barr2−/−) and GRK6 deficient (GRK6−/−) pro-inflammatory macrophages show enhanced migration to chemerin. Control, Barr2−/−, and GRK6−/− pro-inflammatory monocytes/macrophages were fluorescently labeled with calcein and loaded into the upper chamber wells of a Falcon™ HTS FluoroBlok 96-Multiwell Insert System with either 6.25 nM chemerin in the lower chamber to stimulate migration or medium alone as a control. Shown is mean fluorescence intensity (MFI) (measured at 2 minute intervals) of 3–5 independent experiments. Error bars are omitted for clarity. Data was normalized by setting 0 absolute fluorescence as the start point for each curve. The effect of chemerin was examined using an ANCOVA for statistical analysis (Supplemental Figure 1). GRK6−/−macrophage migration was significantly enhanced compared to Barr2−/− and Control (p<0.0001). Barr2−/−macrophage migration was also significant compared to Control (p<0.01).

Article Snippet: Cells were washed and suspended in RPMI 1640 + 1% BSA + 10 mM HEPES, and 1 × 10 5 cells were loaded into FalconTM HTS FluoroBlok 96-Multiwell Insert System chemotaxis chambers (Corning, NY).

Techniques: Migration, Control, Labeling, Fluorescence

Journal: Scientific Reports

Article Title: The oncogenic role of TIMM8A in cancer and the mechanistic insights into the function in breast cancer cells

doi: 10.1038/s41598-025-03331-x

Figure Lengend Snippet: TIMM8A knockdown inhibited the migration and invasion of breast cancer in vitro. ( A – C ) Migration capacity of MCF7 and MDA-MB-231 cells after knockdown of TIMM8A assayed by scratch experiment. Area Recovery (%) = (initial scratch area—final scratch area)/initial scratch area × 100%. ( D – F ) Migration capacity of MCF7 and MDA-MB-231 cells after knockdown of TIMM8A assayed by transwell migration assay. ( G – I ) Invasion capacity of MCF7 and MDA-MB-231 cells after knockdown of TIMM8A assayed by transwell invasion assay. ( J – R ) The expression levels of NF-κB p65 and EMT-related proteins, including α-SMA, Vimentin, E-cadherin and N-cadherin in MCF7 and MDA-MB-231 cells after knockdown of TIMM8A. GAPDH was used as a loading control. The results shown are representative of at least three independent experiments. EMT, epithelial-mesenchymal transition. The student’s t-test or one-way ANOVA was used to detect the differences among groups. * P < 0.05, ** P < 0.01, *** P < 0.001. Samples were derived from the same experiment and gels/blots were processed in parallel. Original blots/gels are presented in the Supplementary File.

Article Snippet: The fundamental procedures were as follows: the transwell chamber (invasion assay: Matrigel® invasion chamber 8.0micron, Corning, USA; migration assay: 24 well multiwell insert System 8.0micron, Falcon, USA) is placed in the 24-well plate, the chamber is called the upper chamber and the plate is called the lower chamber.

Techniques: Knockdown, Migration, In Vitro, Transwell Migration Assay, Transwell Invasion Assay, Expressing, Control, Derivative Assay