primescript tm one step qpcr kit  (tiangen biotech co)

Journal: Frontiers in Cell and Developmental Biology

Article Title: PECAM1 Combines With CXCR4 to Trigger Inflammatory Cell Infiltration and Pulpitis Progression Through Activating the NF-κB Signaling Pathway

doi: 10.3389/fcell.2020.593653

Figure Lengend Snippet: Primer sequences for RT-qPCR.

Article Snippet: Then, qPCR was performed using a one-step PrimeScript TM RT-qPCR kit (Takara Biotechnology Ltd., Dalian, China) on an ABI 7900HT real-time PCR System (Applied Biosystems, Inc., Carlsbad, CA, United States) for RNA quantification.

Techniques:

Journal: Frontiers in Cell and Developmental Biology

Article Title: PECAM1 Combines With CXCR4 to Trigger Inflammatory Cell Infiltration and Pulpitis Progression Through Activating the NF-κB Signaling Pathway

doi: 10.3389/fcell.2020.593653

Figure Lengend Snippet: Knockdown of PECAM1 or CXCR4 alleviates pulpitis in mice. (A) Expression of PECAM1 and CXCR4 in dental pulp tissues determined by RT-qPCR. (B) a diagram presentation of rat treatment: after 1 week of acclimatization, shRNAs of PECAM1 and CXCR1 were injected into the left-side dental pulp tissues of rats. After 48 h, a rat model with pulpitis was induced using LPS. The serum samples were collected 24 h later to examine the expression of inflammatory cytokines in serum. Next, the rats were euthanized through intraperitoneal injection of 150 mg/kg pentobarbital sodium, and then the dental pulp tissues were collected for further use. (C) mRNA expression of PECAM1 and CXCR4 in rat dental pulp tissues examined by RT-qPCR. (D–G) expression of inflammatory cytokines TNF-α, IL-6, IL-8, and VEGF in rat serum determined by ELISA kits. (H) Pathological changes in rat dental pulp tissues observed by HE staining. (I,J) Expression of B-cell-specific protein markers CD19 and CD22 in rat dental pulp tissues determined by IHC staining. In each group, n = 6. Data were collected from three individual experiments and expressed as mean ± SD. Data were analyzed by one-way (D–J) or two-way (A,C) ANOVA followed by Tukey’s multiple comparison test. ** p < 0.01 vs. sham group; ## p < 0.01 vs pulpitis + sh-NC group.

Article Snippet: Then, qPCR was performed using a one-step PrimeScript TM RT-qPCR kit (Takara Biotechnology Ltd., Dalian, China) on an ABI 7900HT real-time PCR System (Applied Biosystems, Inc., Carlsbad, CA, United States) for RNA quantification.

Techniques: Expressing, Quantitative RT-PCR, Injection, Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemistry

Journal: Frontiers in Cell and Developmental Biology

Article Title: PECAM1 Combines With CXCR4 to Trigger Inflammatory Cell Infiltration and Pulpitis Progression Through Activating the NF-κB Signaling Pathway

doi: 10.3389/fcell.2020.593653

Figure Lengend Snippet: PECAM1 and CXCR4 are highly expressed in LPS-treated HDPFs. (A) Morphology of the HDPFs observed under the microscope. (B) Expression of the HDPF-specific biomarkers determined by immunofluorescence staining. (C) Positive expression of HDPF-specific biomarkers examined by flow cytometry. (D–F) mRNA and protein expression of MEF2C. (D) PECAM1 (E) , and CXCR4 (F) in cells examined by RT-qPCR and immunofluorescence staining, respectively. (G–I) , secretion of TNF-α, IL-6, IL-8 in cells determined using ELISA kits. Data were collected from three individual experiments and expressed as mean ± SD. Data were analyzed by unpaired t -test. ** p < 0.01 vs. PBS.

Article Snippet: Then, qPCR was performed using a one-step PrimeScript TM RT-qPCR kit (Takara Biotechnology Ltd., Dalian, China) on an ABI 7900HT real-time PCR System (Applied Biosystems, Inc., Carlsbad, CA, United States) for RNA quantification.

Techniques: Microscopy, Expressing, Immunofluorescence, Staining, Flow Cytometry, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: PECAM1 Combines With CXCR4 to Trigger Inflammatory Cell Infiltration and Pulpitis Progression Through Activating the NF-κB Signaling Pathway

doi: 10.3389/fcell.2020.593653

Figure Lengend Snippet: Downregulation of PECAM1 and CXCR4 suppresses LPS-induced inflammation in HDPFs. (A,B) mRNA (A) and protein (B) expression of PECAM1 and CXCR4 in cells examined by RT-qPCR and western blot assays, respectively. (C–F) Levels of PGE2, TNF-α, IL-6, and IL-8 detected by ELISA kits. (G) Apoptosis of HDPFs determined by the TUNEL assay. Data were collected from three individual experiments and expressed as mean ± SD. Data were analyzed by one-way (C–G) or two-way ANOVA (A,B) followed by Tukey’s multiple comparison test. ** p < 0.01 vs LPS + sh-NC; ## p < 0.01 vs. LPS + oe-NC.

Article Snippet: Then, qPCR was performed using a one-step PrimeScript TM RT-qPCR kit (Takara Biotechnology Ltd., Dalian, China) on an ABI 7900HT real-time PCR System (Applied Biosystems, Inc., Carlsbad, CA, United States) for RNA quantification.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, TUNEL Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: PECAM1 Combines With CXCR4 to Trigger Inflammatory Cell Infiltration and Pulpitis Progression Through Activating the NF-κB Signaling Pathway

doi: 10.3389/fcell.2020.593653

Figure Lengend Snippet: Knockdown of MEF2C reduces LPS-induced inflammation and apoptosis in HDPFs. (A) mRNA expression of MEF2C, PECAM1, and CXCR4 in cells after sh-MEF2C transfection examined by RT-qPCR. (B–E) mRNA and protein levels of inflammatory cytokines PGE2, TNF-α, IL-6, and IL-8 in cells examined by RT-qPCR and ELISA kits. (F) Apoptosis of HDPFs after sh-MEF2C transfection examined by TUNEL assay. Data were collected from three individual experiments and expressed as mean ± SD. Data were analyzed by the unpaired t- test (B–F) or two-way ANOVA (A) followed by Tukey’s multiple comparison test. ** p < 0.01 vs. LPS + sh-NC.

Article Snippet: Then, qPCR was performed using a one-step PrimeScript TM RT-qPCR kit (Takara Biotechnology Ltd., Dalian, China) on an ABI 7900HT real-time PCR System (Applied Biosystems, Inc., Carlsbad, CA, United States) for RNA quantification.

Techniques: Expressing, Transfection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, TUNEL Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: PECAM1 Combines With CXCR4 to Trigger Inflammatory Cell Infiltration and Pulpitis Progression Through Activating the NF-κB Signaling Pathway

doi: 10.3389/fcell.2020.593653

Figure Lengend Snippet: Overexpression of MEF2C reverses the functions of shPECAM1 and shCXCR4 in cells. (A,B) MEF2C overexpression vector was further administrated into HDPFs in the presence of shPECAM1 and shCXCR4. (A,B) mRNA and protein expression of MEF2C, PECAM1, and CXCR4 in cells determined by RT-qPCR and western blot analysis, respectively. (C–F) Levels of PGE2, TNF-α, IL-6, and IL-8 in cells determined by RT-qPCR and ELISA kits, respectively. (G) apoptosis of HDPFs determined by TUNEL assay. Data were collected from three individual experiments and expressed as mean ± SD. Data were analyzed by one-way (C–G) or two-way ANOVA (A,B) followed by Tukey’s multiple comparison test. ** p < 0.01 vs. shPECAM1 + oe-NC; ## p < 0.01 vs. shCXCR4 + oe-NC.

Article Snippet: Then, qPCR was performed using a one-step PrimeScript TM RT-qPCR kit (Takara Biotechnology Ltd., Dalian, China) on an ABI 7900HT real-time PCR System (Applied Biosystems, Inc., Carlsbad, CA, United States) for RNA quantification.

Techniques: Over Expression, Plasmid Preparation, Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, TUNEL Assay