Journal: Cell Death & Disease

Article Title: RASSF1A inhibits PDGFB-driven malignant phenotypes of nasopharyngeal carcinoma cells in a YAP1-dependent manner

doi: 10.1038/s41419-020-03054-z

Figure Lengend Snippet: a A heat map generated using the significantly changed genes categorized in the “cytokine-cytokine receptor interaction pathway” is shown. b , c mRNA expression ( b ) was evaluated by qRT-PCR and protein concentration by ELISA ( c ) in CM of RASSF1A-overexpressing CNE-2 cells, RASSF1A-depleted CNE-1 cells and their corresponding control cells, The data are presented as the mean ± S.D. values, ** p < 0.01, Student’s t test. d – g PDGFB was transiently knocked down with a pool of siRNA or treated with a neutralizing antibody for PDGF-BB (10 µg/mL) in RASSF1A-depleted CNE-1 cells. PDGF-BB secretion in the CM was measured by ELISA ( d ), ** p < 0.01, Student’s t test. e Number of spheroids formed was determined via microscopy, and representative images ( e left panel) are shown. The formed spheroids were compared ( e right panel), the data are presented as the mean ± S.D. values, * p < 0.05, ** p < 0.01, Student’s t test; ns: non-sinificant. Scale bar: 200 µm. Representative images of the migration assay ( f ) and invasion assay ( g ) are shown, the data are presented as the mean ± S.D. values, ** p < 0.01, Student’s t test. Scale bar: 100 µm. h – j Recombinant PDGF-BB or IgG was added to RASSF1A-overexpressing CNE-2 cells. Representative images of sphere formation ( h ) (Scale bar: 200 µm.), migration ( i ) and invasion ( j ) assays (Scale bar: 100 µm) of RASSF1A-overexpressing CNE-2 cells treated with PDGF-BB (the culture medium was supplemented with 20 ng/ml or an equal volume of control IgG) are shown, The data are presented as the mean ± S.D. values, * p < 0.05, ** p < 0.01, Student’s t test.

Article Snippet: Cells were plated in 6-well plates (Corning, USA) and treated with humane recombinant PDGF-BB (220-BB-010, R&D Systems, USA) or Immunoglobulin G (IgG) control (AB-108-C, R&D Systems, USA) or neutralizing antibody against PDGF-BB (AB-220-NA, R&D Systems, USA) or latrunculin b (LTB, ab144291, Abcam, UK) 12 h after plating.

Techniques: Generated, Expressing, Quantitative RT-PCR, Protein Concentration, Enzyme-linked Immunosorbent Assay, Control, Microscopy, Migration, Invasion Assay, Recombinant

Journal: Cell Death & Disease

Article Title: RASSF1A inhibits PDGFB-driven malignant phenotypes of nasopharyngeal carcinoma cells in a YAP1-dependent manner

doi: 10.1038/s41419-020-03054-z

Figure Lengend Snippet: a – c YAP1 was transiently knocked down in RASSF1A-depleted CNE-1 cells. a In the indicated cells, YAP1 protein expression was assessed by using western blotting; b PDGFB, CYP61 and CTGF mRNA expression was assessed by qRT-PCR; c Concentration of PDGF-BB secreted in CM was measured by ELISA. The data are presented as the mean ± S.D. values, * p < 0.05, ** p < 0.01, Student’s t test. d – f Recombinant PDGF-BB or IgG was added to YAP1-silenced NPC cells. d The formed spheroids were counted via microscopy, and ( e ) representative images are shown. f The impact of PDGF-BB treatment on the migration and invasion of RASSF1A-depleted cells was determined by Transwell assays. The data are presented as the mean ± S.D. values, * p < 0.05, ** p < 0.01, Student’s t test. Scale bar: 200 µm.

Article Snippet: Cells were plated in 6-well plates (Corning, USA) and treated with humane recombinant PDGF-BB (220-BB-010, R&D Systems, USA) or Immunoglobulin G (IgG) control (AB-108-C, R&D Systems, USA) or neutralizing antibody against PDGF-BB (AB-220-NA, R&D Systems, USA) or latrunculin b (LTB, ab144291, Abcam, UK) 12 h after plating.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Concentration Assay, Enzyme-linked Immunosorbent Assay, Recombinant, Microscopy, Migration