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Fig. 3 MPM upregulated miR-205 and downregulated <t>PAX9</t> in KD cells. (A) Quantitative RT-PCR of miR-27b-3p, miR- 27b-5p, miR-133b, miR-205-3p, miR-205-5p, miR-497-5p, and miR-655-3p after treatment of KD cells with 1 μM MPM for 72 h. Data are presented as the mean ± SD. *P < 0.05 (n = 3). (B) Quantitative RT-PCR for PAX9 and RARA after treatment of KD cells with 1 μM MPM for 72 h. Data are presented as the mean ± SD. *P < 0.05 versus control (n = 4). (C) Western blotting of KD cells treated with 1 μM MPM for 72 h. β-ACTIN was served as an internal control.
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Fig. 3 MPM upregulated miR-205 and downregulated PAX9 in KD cells. (A) Quantitative RT-PCR of miR-27b-3p, miR- 27b-5p, miR-133b, miR-205-3p, miR-205-5p, miR-497-5p, and miR-655-3p after treatment of KD cells with 1 μM MPM for 72 h. Data are presented as the mean ± SD. *P < 0.05 (n = 3). (B) Quantitative RT-PCR for PAX9 and RARA after treatment of KD cells with 1 μM MPM for 72 h. Data are presented as the mean ± SD. *P < 0.05 versus control (n = 4). (C) Western blotting of KD cells treated with 1 μM MPM for 72 h. β-ACTIN was served as an internal control.

Journal: Biomedical research (Tokyo, Japan)

Article Title: Mycophenolate mofetil reduces cell viability associated with the miR-205-PAX9 pathway in human lip fibroblast cells.

doi: 10.2220/biomedres.46.1

Figure Lengend Snippet: Fig. 3 MPM upregulated miR-205 and downregulated PAX9 in KD cells. (A) Quantitative RT-PCR of miR-27b-3p, miR- 27b-5p, miR-133b, miR-205-3p, miR-205-5p, miR-497-5p, and miR-655-3p after treatment of KD cells with 1 μM MPM for 72 h. Data are presented as the mean ± SD. *P < 0.05 (n = 3). (B) Quantitative RT-PCR for PAX9 and RARA after treatment of KD cells with 1 μM MPM for 72 h. Data are presented as the mean ± SD. *P < 0.05 versus control (n = 4). (C) Western blotting of KD cells treated with 1 μM MPM for 72 h. β-ACTIN was served as an internal control.

Article Snippet: Supplementary Table S1 Antibody lists for western blotting Antibody name Vendor Catalog number Concentration β-ACTIN Medical & Biological Laboratories M177-3 1 : 3000 Cleaved CASPASE-3 Cell Signaling Technology 9661 1 : 3000 CCND1 Santa Cruz Biotechnology sc-8396 1 : 500 CCNE Santa Cruz Biotechnology sc-377100 1 : 1000 CDK2 Santa Cruz Biotechnology sc-6248 1 : 1000 CDK4 Santa Cruz Biotechnology sc-56277 1 : 1000 CDK6 Santa Cruz Biotechnology sc-53638 1 : 500 PAX9 Santa Cruz Biotechnology sc-56823 1 : 500 RARA Santa Cruz Biotechnology sc-515796 1 : 500 Rabbit IgG HRP Cell Signaling Technology 7074 1 : 10000 Mouse IgG HRP Cell Signaling Technology 7976 1 : 10000 Supplementary Fig. S2 Putative target site for miR-205-3p and miR-205-5p in the PAX9 and RARA 3’ UTR.

Techniques: Quantitative RT-PCR, Control, Western Blot

Fig. 4 Overexpression of PAX9 suppressed MPM-induced cell proliferation inhibition in KD cells. (A) Quantitative RT-PCR analysis of PAX9 expression after transfected KD cells with 600 ng pcDNA3.1-neo-PAX9 for 24 h. Data are presented as the mean ± SD. ***P < 0.001 (n = 4). (B) Western blotting of KD cells transfected with 600 ng pcDNA3.1-neo-PAX9 for 24 h. β-ACTIN was served as an internal control. (C) Proliferation of KD cells treated with 1 μM MPM and/or 600 ng pcDNA3.1- neo-PAX9 for 72 h. Data are presented as the mean ± SD. *P < 0.05, **P < 0.01 and ***P < 0.001 (n = 6). (D) BrdU staining (green) of KD cells after treatment with 1 μM MPM and/or 600 ng pcDNA3.1-neo-PAX9 for 72 h. BrdU positive cells were stained in green. The nuclei were counterstained with Hoechst 33342 (blue). The arrows showed BrdU positive nuclei. Scale bar, 50 μm. Graph shows the quantification of BrdU positive cells. Data are presented as the mean ± SD. *P < 0.05, **P < 0.01 versus control (n = 6).

Journal: Biomedical research (Tokyo, Japan)

Article Title: Mycophenolate mofetil reduces cell viability associated with the miR-205-PAX9 pathway in human lip fibroblast cells.

doi: 10.2220/biomedres.46.1

Figure Lengend Snippet: Fig. 4 Overexpression of PAX9 suppressed MPM-induced cell proliferation inhibition in KD cells. (A) Quantitative RT-PCR analysis of PAX9 expression after transfected KD cells with 600 ng pcDNA3.1-neo-PAX9 for 24 h. Data are presented as the mean ± SD. ***P < 0.001 (n = 4). (B) Western blotting of KD cells transfected with 600 ng pcDNA3.1-neo-PAX9 for 24 h. β-ACTIN was served as an internal control. (C) Proliferation of KD cells treated with 1 μM MPM and/or 600 ng pcDNA3.1- neo-PAX9 for 72 h. Data are presented as the mean ± SD. *P < 0.05, **P < 0.01 and ***P < 0.001 (n = 6). (D) BrdU staining (green) of KD cells after treatment with 1 μM MPM and/or 600 ng pcDNA3.1-neo-PAX9 for 72 h. BrdU positive cells were stained in green. The nuclei were counterstained with Hoechst 33342 (blue). The arrows showed BrdU positive nuclei. Scale bar, 50 μm. Graph shows the quantification of BrdU positive cells. Data are presented as the mean ± SD. *P < 0.05, **P < 0.01 versus control (n = 6).

Article Snippet: Supplementary Table S1 Antibody lists for western blotting Antibody name Vendor Catalog number Concentration β-ACTIN Medical & Biological Laboratories M177-3 1 : 3000 Cleaved CASPASE-3 Cell Signaling Technology 9661 1 : 3000 CCND1 Santa Cruz Biotechnology sc-8396 1 : 500 CCNE Santa Cruz Biotechnology sc-377100 1 : 1000 CDK2 Santa Cruz Biotechnology sc-6248 1 : 1000 CDK4 Santa Cruz Biotechnology sc-56277 1 : 1000 CDK6 Santa Cruz Biotechnology sc-53638 1 : 500 PAX9 Santa Cruz Biotechnology sc-56823 1 : 500 RARA Santa Cruz Biotechnology sc-515796 1 : 500 Rabbit IgG HRP Cell Signaling Technology 7074 1 : 10000 Mouse IgG HRP Cell Signaling Technology 7976 1 : 10000 Supplementary Fig. S2 Putative target site for miR-205-3p and miR-205-5p in the PAX9 and RARA 3’ UTR.

Techniques: Over Expression, Inhibition, Quantitative RT-PCR, Expressing, Transfection, Western Blot, Control, BrdU Staining, Staining