Journal: Molecules

Article Title: RORα Regulates Odontoblastic Differentiation and Mediates the Pro-Odontogenic Effect of Melatonin on Dental Papilla Cells

doi: 10.3390/molecules26041098

Figure Lengend Snippet: Figure 1. Expression of nuclear receptor retinoid acid receptor-related orphan receptors (RORs) in rat dental papilla cells (rDPCs). (a) Total RNA was extracted from rDPCs and detected by reverse transcription polymerase chain reaction (RT-PCR) and agarose gel electrophoresis assay using specific primers for RORα, RORβ, RORγ, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). RT-PCR without primers served as the negative control (NC); (b) Immunofluorescence staining was performed with polyclonal anti-RORα antibody (green), and nuclei were labelled with 4-6-diamidino-2-phenylindole (DAPI, blue). Scale bar: 50 µm.

Article Snippet: Subsequently, they were incubated overnight at 4 ◦C with the following primary antibodies: polyclonal rabbit anti-RORα antibody (1:2000, PA5-23268, Thermo Scientific, MA, USA), monoclonal mouse anti-DSPP antibody (1:500, sc-73632, Santa Cruz, CA, USA), polyclonal rabbit anti-DMP1 antibody (1:1000, NBP 1-45525, Novus Biologicals, Littleton, CO, USA), and monoclonal mouse anti-β-actin (1:1000, AF0003, Beyotime, Shanghai, China).

Techniques: Expressing, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Negative Control, Staining

Journal: Molecules

Article Title: RORα Regulates Odontoblastic Differentiation and Mediates the Pro-Odontogenic Effect of Melatonin on Dental Papilla Cells

doi: 10.3390/molecules26041098

Figure Lengend Snippet: Figure 2. The expression pattern of retinoid acid receptor-related orphan receptor α (RORα) and odontogenic markers in rat dental papilla cells (rDPCs) during odontoblastic differentiation. rDPCs were cultured in maintained medium (control) or odontogenic induction medium (OS) for 3 and 7 days (3d and 7d), respectively. (a) The mRNA levels of RORα, dentin sialophosphoprotein (DSPP), dentin matrix protein 1 (DMP1), and alkaline phosphatase (ALP) were quantified by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Glyceraldehyde-3-phosphate dehydroge- nase (GAPDH) was used as the normalisation control; (b) The protein levels of RORα, DSPP, and DMP1 were determined by western blotting and normalised to the protein level of β-actin. All data are presented as the mean ± SD (n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Control group.

Article Snippet: Subsequently, they were incubated overnight at 4 ◦C with the following primary antibodies: polyclonal rabbit anti-RORα antibody (1:2000, PA5-23268, Thermo Scientific, MA, USA), monoclonal mouse anti-DSPP antibody (1:500, sc-73632, Santa Cruz, CA, USA), polyclonal rabbit anti-DMP1 antibody (1:1000, NBP 1-45525, Novus Biologicals, Littleton, CO, USA), and monoclonal mouse anti-β-actin (1:1000, AF0003, Beyotime, Shanghai, China).

Techniques: Expressing, Cell Culture, Control, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot

Journal: Molecules

Article Title: RORα Regulates Odontoblastic Differentiation and Mediates the Pro-Odontogenic Effect of Melatonin on Dental Papilla Cells

doi: 10.3390/molecules26041098

Figure Lengend Snippet: Figure 4. Effect of retinoid acid receptor-related orphan receptor α (RORα) overexpression on odontoblastic differentiation of rat dental papilla cells (rDPCs). rDPCs were transfected with pcDNA3.1-RORα (RORα group) or pcDNA3.1-NC (negative control group) for 24 h and then cultured in control or odontogenic induction (OS) medium for 3 or 7 days. (a) The transfection efficiency of RORα overexpression was assessed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blotting. (b) The mRNA levels of dentin sialophosphoprotein (DSPP), dentin matrix protein 1 (DMP1), and alkaline phosphatase (ALP) were detected by qRT-PCR after 3-day induction. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the normalisation control; (c) The protein levels of DSPP and DMP1 were measured by western blotting after 7-day induction. β-actin was used as the internal control; (d) ALP activity in cellular lysates was determined after 7-day odontogenic induction. (e) The formation of mineralized nodules was visualized by alizarin red staining at 7 days after induction. Scale bar: 100 µm. All data are presented as the mean ± SD (n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. pcDNA3.1-NC or Control-NC group, # p < 0.05, ### p < 0.001 vs. OS-NC group.

Article Snippet: Subsequently, they were incubated overnight at 4 ◦C with the following primary antibodies: polyclonal rabbit anti-RORα antibody (1:2000, PA5-23268, Thermo Scientific, MA, USA), monoclonal mouse anti-DSPP antibody (1:500, sc-73632, Santa Cruz, CA, USA), polyclonal rabbit anti-DMP1 antibody (1:1000, NBP 1-45525, Novus Biologicals, Littleton, CO, USA), and monoclonal mouse anti-β-actin (1:1000, AF0003, Beyotime, Shanghai, China).

Techniques: Over Expression, Transfection, Negative Control, Cell Culture, Control, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Activity Assay, Staining

Journal: Molecules

Article Title: RORα Regulates Odontoblastic Differentiation and Mediates the Pro-Odontogenic Effect of Melatonin on Dental Papilla Cells

doi: 10.3390/molecules26041098

Figure Lengend Snippet: Figure 5. Effect of retinoid acid receptor-related orphan receptor α (RORα) knockdown on odontoblastic differentiation of rat dental papilla cells (rDPCs). rDPCs were transfected with small interfering (si) RORα or si negative control (NC) for 8 h and then cultured in control or odontogenic induction (OS) medium for 3 or 7 days. (a) The knockdown efficiency of RORα was examined by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blotting; (b) The mRNA levels of dentin sialophosphoprotein (DSPP), dentin matrix protein 1 (DMP1), and alkaline phosphatase (ALP) were measured by qRT-PCR after 3-day induction. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the internal control; (c) The protein levels of DSPP and DMP1 were detected by western blotting after 7-day induction and normalised to β-actin levels; (d) ALP activity was analysed after 7 days of odontogenic induction. (e) The formation of mineralized nodules was assessed by alizarin red staining at day 7. Scale bar: 100 µm. All data are presented as the mean ± SD (n = 3). ** p < 0.01, *** p < 0.001 vs. si NC or Control-si NC group, # p < 0.05, ## p < 0.01 vs. OS-si NC group.

Article Snippet: Subsequently, they were incubated overnight at 4 ◦C with the following primary antibodies: polyclonal rabbit anti-RORα antibody (1:2000, PA5-23268, Thermo Scientific, MA, USA), monoclonal mouse anti-DSPP antibody (1:500, sc-73632, Santa Cruz, CA, USA), polyclonal rabbit anti-DMP1 antibody (1:1000, NBP 1-45525, Novus Biologicals, Littleton, CO, USA), and monoclonal mouse anti-β-actin (1:1000, AF0003, Beyotime, Shanghai, China).

Techniques: Knockdown, Transfection, Negative Control, Cell Culture, Control, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Activity Assay, Staining

Journal: eLife

Article Title: Hsf1 and the molecular chaperone Hsp90 support a ‘rewiring stress response’ leading to an adaptive cell size increase in chronic stress

doi: 10.7554/eLife.88658

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-Hsp25/27 (Mouse monoclonal) , StressMarq , SMC-114 , 1:1000.

Techniques: Generated, Recombinant, Plasmid Preparation, Over Expression, Mutagenesis, Control, Sequencing, shRNA, Flow Cytometry