Journal: PLOS ONE

Article Title: Cyclosporine A causes gingival overgrowth via reduced G 1 cell cycle arrest in gingival fibroblasts

doi: 10.1371/journal.pone.0309189

Figure Lengend Snippet: Primers used for reverse transcription-quantitative PCR.

Article Snippet: The rabbit anti-CDC25A antibody (cat. no. 55031-1-AP) was purchased from Proteintech Group, Inc (Rosemont, IL, USA).

Techniques: Sequencing

Journal: PLOS ONE

Article Title: Cyclosporine A causes gingival overgrowth via reduced G 1 cell cycle arrest in gingival fibroblasts

doi: 10.1371/journal.pone.0309189

Figure Lengend Snippet: Semiconfluent cells were incubated in D-MEM containing 2% serum with or without (control) 200 ng/mL (166 nM) CsA for 12 h, after which reverse transcription-quantitative PCR analysis was performed. Relative quantification was performed using the 2‑ΔΔCq method. After normalization to GAPDH, RNA ratios in treated vs. control cultures were determined. Data are presented as means ± SEM. * P <0.05 compared with the control using Welch’s t-test (n = 3). (A) G 1 -S transition-promoted genes ( CDC25A , cell division cycle 25A; CDK2 , cyclin dependent kinase 2; CYCLIN E1 , E-type cyclin; MYC , MYC proto-oncogene, bHLH transcription factor). (B) G 1 arrest-induced genes ( ATM , ATM serine/threonine kinase; ATR , ATR serine/threonine kinase; P21 , cyclin dependent kinase inhibitor 1A; P27 , cyclin dependent kinase inhibitor 1B; P53 , tumor protein p53; RB1 , RB transcriptional corepressor 1; SMAD3 , SMAD family member 3; SMAD4 , SMAD family member 4).

Article Snippet: The rabbit anti-CDC25A antibody (cat. no. 55031-1-AP) was purchased from Proteintech Group, Inc (Rosemont, IL, USA).

Techniques: Incubation, Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative Proteomics

Journal: PLOS ONE

Article Title: Cyclosporine A causes gingival overgrowth via reduced G 1 cell cycle arrest in gingival fibroblasts

doi: 10.1371/journal.pone.0309189

Figure Lengend Snippet: Semiconfluent cells were incubated in D-MEM containing 2% serum with or without (control) 200 ng/mL (166 nM) CsA for 24 h and were then assessed using western blotting, after which the fold change compared with the control was determined. The band images shown are representative of results from three independent experiments. Data are presented as means ± SEM. * P <0.05 compared with the control using Welch’s t-test (n = 3). (A) G 1 -S transition-promoted proteins (CDC25A, cell division cycle 25A; CYCLIN E1, E-type cyclin; pCDK2, phospho-cyclin dependent kinase 2; RB1, RB transcriptional corepressor 1). (B) G 1 arrest-induced proteins (P21, cyclin dependent kinase inhibitor 1A; SMAD3, SMAD family member 3; SMAD4, SMAD family member 4).

Article Snippet: The rabbit anti-CDC25A antibody (cat. no. 55031-1-AP) was purchased from Proteintech Group, Inc (Rosemont, IL, USA).

Techniques: Incubation, Control, Western Blot

Journal: PLOS ONE

Article Title: Cyclosporine A causes gingival overgrowth via reduced G 1 cell cycle arrest in gingival fibroblasts

doi: 10.1371/journal.pone.0309189

Figure Lengend Snippet: CsA induces the downregulation of SMAD3 and SMAD4, which leads to a decrease in P21. CsA also induces the upregulation of CDC25A, CYCLIN E, phosphorylated CDK2 and phosphorylated RB, which results in the inhibition of G 1 cell cycle arrest. Light blue (factors of G 1 arrest-inducing signaling) and light green (factors of G 1 /S transition-inducing signaling) rectangles similarly denote the molecules analyzed in this study. The blue or red large arrows denote downregulation or upregulation, respectively, following CsA treatment. ATM, ATM serine/threonine kinase; ATR, ATR serine/threonine kinase; CDC25A, cell division cycle 25A; CYCLIN E, E-type cyclin; MYC, MYC proto-oncogene, bHLH transcription factor; pCDK2, phosphorylated cyclin dependent kinase 2; pRB1, phosphorylated RB transcriptional corepressor 1; P21, cyclin dependent kinase inhibitor 1A; P27, cyclin dependent kinase inhibitor 1B; P53, tumor protein p53; RB1, RB transcriptional corepressor 1; SMAD3, SMAD family member 3; SMAD4, SMAD family member 4.

Article Snippet: The rabbit anti-CDC25A antibody (cat. no. 55031-1-AP) was purchased from Proteintech Group, Inc (Rosemont, IL, USA).

Techniques: Inhibition