Journal: Cell Death & Disease

Article Title: Colorectal cancer cell intrinsic fibroblast activation protein alpha binds to Enolase1 and activates NF-κB pathway to promote metastasis

doi: 10.1038/s41419-021-03823-4

Figure Lengend Snippet: A Immunofluorescence analysis of FAP location in HCT116-FAP and HCT8-FAP cells. Error bar = 30 μm. B ELISA assay detects the concentration of FAP in media of HCT116-FAP, HCT8-FAP, HCT116-Ctrl, and HCT8-Ctrl cells. Ctrl: control; FAP: FAP overexpression. C Migration assays of HCT116 and HCT8 cells treated with control medium and FAP-overexpressed medium. Error bar = 100 μm. * p < 0.05, *** p < 0.001. The CM was added into the upper chamber. CM: conditioned medium. D Invasion assays of HCT116 and HCT8 cells treated with control medium and FAP-overexpressed medium. Error bar = 100 μm. * p < 0.05, *** p < 0.001. The CM was added into the upper chamber. E Migration assays of HCT116 and HCT8 cells treated with different concentration of recombinant FAP. Error bar = 100 μm. ** p < 0.01, *** p < 0.001. F Invasion assays of HCT116 and HCT8 cells treated with different concentration of recombinant FAP. Error bar = 100 μm. ** p < 0.01, *** p < 0.001.

Article Snippet: FAP concentration was detected using FAP DuoSet ELISA kit (R&D Systems, DY3715) according to the manufacturer’s protocol.

Techniques: Immunofluorescence, Enzyme-linked Immunosorbent Assay, Concentration Assay, Control, Over Expression, Migration, Recombinant

Journal: Current Issues in Molecular Biology

Article Title: Oncological Safety of High Hydrostatic Pressure Treatment: Effects on Cancer-Associated Fibroblast-like Transdifferentiation of Adipose Stromal Cells

doi: 10.3390/cimb48010091

Figure Lengend Snippet: Protein expression analysis of cancer-associated fibroblast (CAF) markers in stimulated human adipose stromal cells (hASCs). HASCs were incubated for 96 h with tumor-conditioned medium (TCM), 0 MPa-CM, or 300 MPa-CM derived from HNSCC16 or UTSCC14 cells, TGF-β1 (10 ng/mL), or serum-free control medium, respectively. ( A ) α-SMA protein expression was determined by Western blot analysis. Representative Western blot membrane with protein bands of α-SMA following incubation with primary and secondary antibody is shown. Precision Plus Protein™ All Blue prestained standard served as molecular weight marker. Densitometric quantification of α-SMA protein expression was evaluated and total protein normalization was performed using Bio-Rad Stain-Free™ technology. Data represents n = 4 independent donors (mean ± SD). Results are presented as relative protein expression normalized to the serum-free control (set to 1; illustrated as dashed lines). ( B ) FAP levels were determined by ELISA in hASCs from six independent donors after 96 h of stimulation with the respective conditioned media. Results are presented as relative protein expression normalized to the serum-free control (set to 1; illustrated as dashed lines). Statistical significance was assessed using predefined post hoc comparisons following one-way ANOVA or Kruskal–Wallis testing. Differences were evaluated relative to the control group or between specific treatment conditions as indicated in the figures. Statistically significant differences are indicated by asterisks (* p < 0.05, ** p < 0.01).

Article Snippet: Determination of FAP concentration was performed on cell lysates using the DuoSet Human FAP enzyme-linked immunosorbent assay (ELISA) kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Expressing, Incubation, Derivative Assay, Control, Western Blot, Membrane, Molecular Weight, Marker, Staining, Enzyme-linked Immunosorbent Assay