Journal: Cancer Biology & Medicine

Article Title: Systematic screening reveals synergistic interactions that overcome MAPK inhibitor resistance in cancer cells

doi: 10.20892/j.issn.2095-3941.2020.0560

Figure Lengend Snippet: The disruption of both HDGF and LGR5 synergistically represses the emergence of BRAFi-responsive oncogenic signals. (A) Representative immunofluorescence micrographs showing KLF4 (red) and SOX2 (green) expressions, β-catenin (red), and CD133 (green) expressions, and nestin (red) and phospho-FAK (green) expressions in cells treated with dimethyl sulfoxide (DMSO), vemurafenib, vemurafenib + siLGR5 , vemurafenib + siHDGF , siLGR5 + siHDGF , or vemurafenib + siLGR5 + siHDGF . Scale bars, 10 μm (white). (B) Western blot showing OCT4, NGFR, nestin, BMI1, and CD133 levels in DMSO-, vemurafenib-, vemurafenib + siLGR5 -, vemurafenib + siHDGF -, siLGR5 + siHDGF -, and vemurafenib + siLGR5 + siHDGF -treated A375 cell samples. (C) Western blot showing TGFβ3 and phospho-SMAD2 (Ser250) levels in vemurafenib + siHDGF + siLGR5 -, siHDGF + siLGR5 -, vemurafenib-, and DMSO-treated A375 or SK-MEL-28 cell samples. (D) A proposed model for adaptive HDGF-LGR5 drug resistance to MAPK inhibitors.

Article Snippet: Primary antibodies were anti-mouse-SOX2 (Sino Biological), anti-rabbit-KLF4 (Beyotime Biotechnology), anti-rabbit-Phospho-FAK (Beyotime Biotechnology), and anti-nestin (Beyotime Biotechnology).

Techniques: Immunofluorescence, Western Blot

Journal: International Journal of Biological Sciences

Article Title: EXOSC5 maintains cancer stem cell activity in endometrial cancer by regulating the NTN4/integrin β1 signalling axis

doi: 10.7150/ijbs.86275

Figure Lengend Snippet: Secreted Netrin4 upregulates c-MYC by triggering integrin β1/FAK/β-catenin signalling axis. (A, B) Serum-starved AN3CA (A) and HEC1A (B) cells were treated with 50 ng/ml rhNTN4 and harvested at the indicated time points. Activation of FAK, SRC, and β-catenin was analysed using Western blot. (C) AN3CA cells, treated with 50 ng/ml rhNTN4 for 1 hour, had their FAK phosphorylation at Tyr397 visualized using immunofluorescence staining. DAPI was utilized for nuclear counterstaining. (D, E) AN3CA cells were treated with 50 ng/ml rhNTN4 or 20 µM Wnt agonist for 16 hours. β-catenin expression was then visualized using immunofluorescence staining (D). Scale bars represented 100 µm in length. The dotted circles in the magnified images highlight the cell nuclei based on DAPI signals. ImageJ was used to quantify the nuclear/cytoplasmic intensities of β-catenin. The ratios of nuclear/cytoplasmic β-catenin, determined from two objective fields, are presented as mean ± SD (E). *, p< 0.05; **, p< 0.01 by Student's t -test when compared to untreated control cells (ctrl). (F) AN3CA and EMC6 cells, treated with control IgG or 1 µg/ml anti-CD29 for 16 hours, were assessed for FAK phosphorylation and c-MYC expression using Western blot. (G) AN3CA and EMC6 cells, treated with either 0.01% DMSO or 1 µM Defactinib for 16 hours, were analyzed for FAK phosphorylation and c-MYC expression via Western blot. (H, I) The protein expression of p-FAK Tyr397 and c-MYC in EC tissues was visualized using IHC staining on tissue microarray slides. Representative images showcasing high/low staining intensities of p-FAK Tyr397 and c-MYC are provided in (H). Their correlation was then evaluated using Pearson's correlation analysis (I).

Article Snippet: Catalog No. GTX118473), anti-NTN4 (Sigma-Aldrich, Catalog No. HPA049832), anti-phospho-FAK Tyr397 (ABclonal, Inc., Woburn, MA, USA.

Techniques: Activation Assay, Western Blot, Immunofluorescence, Staining, Expressing, Immunohistochemistry, Microarray