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

Journal: ACS Omega

Article Title: In Vitro Enhanced Performance of Human Platelet Lysate Gel Integrated with Mesoporous Silica Nanoparticle/Carboxymethyl Chitosan Composite Hydrogel: Structural Stability and Biological Activities for Chronic Wound Healing

doi: 10.1021/acsomega.5c13494

Figure Lengend Snippet: Measurements of total protein and growth factors released from hPLG and xPMC/hPLG. (A) Cumulative release percentage of total protein over a 4 day period, determined by BCA assay. (B) SDS-PAGE analysis results of the PBS supernatants collected over a 4 day period, stained with Coomassie blue. Cumulative release of (C) TGF-β1 and (D) PDGF-BB over a 3 day period. (E) Total protein contents found in the PBS supernatants of hPLG and hPLG combined with either PM or xCMC, assessed after a 2 day incubation. Data are expressed as mean ± SD ( n = 3). * p < 0.05, ** p < 0.01. Statistical significance in (C,D) was analyzed at the same time point.

Article Snippet: To unequivocally determine the role of PDGF-BB specifically as a primary GF mediating hPL-induced periodontal fibroblast migration, PDGF-BB activity was inhibited by preincubating xPMC/hPLG spots with 0.2 μg/mL of a PDGF-BB neutralizing antibody (AF-220-NA, R&D Systems) for 45 min. As a negative control, xPMC/hPLG spots were pretreated for 45 min with an isotype control goat IgG antibody (0.2 μg/mL; AB-108 C, R&D Systems).

Techniques: BIA-KA, SDS Page, Staining, Incubation

Journal: ACS Omega

Article Title: In Vitro Enhanced Performance of Human Platelet Lysate Gel Integrated with Mesoporous Silica Nanoparticle/Carboxymethyl Chitosan Composite Hydrogel: Structural Stability and Biological Activities for Chronic Wound Healing

doi: 10.1021/acsomega.5c13494

Figure Lengend Snippet: Effect of xPMC/hPLG on the migration of periodontal fibroblasts. (A) Cells were seeded and exposed to chemokines from different gel spots for 6 h. The migratory cells surrounding the edge of gel spots of (i) hPPG, (ii) hPLG, (iii) xPMC/hPLG, (iv) xPMC/hPLG pretreated with IgG isotype control antibody (Ab), and (v) xPMC/hPLG pretreated with neutralizing antibody (nAb) against PDGF-BB were stained with crystal violet (A), and the cell density (cells/mm 2 ) was quantified from the number of cells that migrated close to each gel spot within a 500 μm radial distance from the gel edge (B). The results are expressed as mean ± SD ( n = 8). Different letters indicate significant differences ( p < 0.05).

Article Snippet: To unequivocally determine the role of PDGF-BB specifically as a primary GF mediating hPL-induced periodontal fibroblast migration, PDGF-BB activity was inhibited by preincubating xPMC/hPLG spots with 0.2 μg/mL of a PDGF-BB neutralizing antibody (AF-220-NA, R&D Systems) for 45 min. As a negative control, xPMC/hPLG spots were pretreated for 45 min with an isotype control goat IgG antibody (0.2 μg/mL; AB-108 C, R&D Systems).

Techniques: Migration, Control, Staining

Journal: ACS Omega

Article Title: In Vitro Enhanced Performance of Human Platelet Lysate Gel Integrated with Mesoporous Silica Nanoparticle/Carboxymethyl Chitosan Composite Hydrogel: Structural Stability and Biological Activities for Chronic Wound Healing

doi: 10.1021/acsomega.5c13494

Figure Lengend Snippet: Schematic image of the proposed concentration-dependent biphasic effect of xPMC/hPLG on the chemotaxis and cell doubling of periodontal fibroblasts. Color gradient represents a concentration gradient from high (red) to low (blue) concentrations of hPL-derived mediators released from the xPMC/hPLG hydrogel. The hydrogel gave rise to a concentration gradient of a GF, i.e., PDGF-BB, declining with increasing distance away from the hydrogel. Fibroblasts in Area 2 were exposed to a low concentration of PDGF-BB sufficient to induce their migration toward the hydrogel. As the cells ascended the concentration gradient, the signal for chemotaxis was switched off, but the signal for cell division was switched on, thereby resulting in the presence of proliferating cells at Area 1, adjacent to the hydrogel.

Article Snippet: To unequivocally determine the role of PDGF-BB specifically as a primary GF mediating hPL-induced periodontal fibroblast migration, PDGF-BB activity was inhibited by preincubating xPMC/hPLG spots with 0.2 μg/mL of a PDGF-BB neutralizing antibody (AF-220-NA, R&D Systems) for 45 min. As a negative control, xPMC/hPLG spots were pretreated for 45 min with an isotype control goat IgG antibody (0.2 μg/mL; AB-108 C, R&D Systems).

Techniques: Concentration Assay, Chemotaxis Assay, Derivative Assay, Migration

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: Pericytes contribute to airway remodeling in a mouse model of chronic allergic asthma

doi: 10.1152/ajplung.00286.2014

Figure Lengend Snippet: A : qPCR analysis of PDGF-BB and PDGFRβ mRNA expression in whole-lung lysates from saline and HDM-exposed mice (i.n. 5 days/wk for 5 wk); n = 5 representative of 2 independent experiments. ELISA analysis of PDGF-BB expression in BAL supernatants ( B ) and immunoblotting for PDGF-BB protein expression ( C ) in whole-lung lysates from saline and HDM-exposed mice (i.n. 5 days/wk for 5 wk); n = 5–6 (ELISA) and n = 3 (immunoblotting) per group, representative of 2 independent experiments. The immunoblot shown in C presents representative lanes for saline and HDM-exposed mice from 3 replicates per group run on the same gel. D : quantification of PDGF-BB expression assessed by immunoblotting in whole lung homogenates from saline and HDM-exposed mice. * P < 0.05, ** P < 0.01.

Article Snippet: The membranes were incubated in blocking buffer (5% skim milk) in Tris-buffered saline containing 0.5% Tween for 1 h at room temperature and probed with a polyclonal rabbit anti-human PDGF-BB antibody (Aviva Systems Biology, San Diego, CA) diluted 1:1,000 in blocking buffer.

Techniques: Expressing, Saline, Enzyme-linked Immunosorbent Assay, Western Blot