Real-time quantitative PCR (qPCR) was performed using a Genomic DNA Purification Kit (Tiangen, China). The Ct values obtained from different samples were compared using the 2−ΔΔCt method. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as an internal reference transcript. The primers ...
qPCR was performed following the manufacturer's instructions. The following sequences were used for qPCR primer: DCLK1 (sense, 5′-AGCCTC ATGCGGTTAATTTG-3′; anti-sense, 5′-TCTTTGAAT TAGCGCCTGGT-3′), YAP (sense, 5′-CAGAACCGT...
The cDNA products were diluted to 40 µl with sterilized ddH2O.1 µl (12.5 µg) cDNA was used for the real-time PCR reaction using 2xSG Fast qPCR Master Mix. Primers and PCR operation were according to 1.2. Real-time PCR of PTLCs PTLCs were collected and total RNA was ...
Finally, the specific sequences from immunoprecipitated and input DNA were determined by real-time quantitative PCR (qPCR) for the upstream of IL-8 and SRGN promoter regions. The four primer pairs for the IL-8 and SRGN promoter regions used in qPCR analyses are listed in supplementary Table s...
The amount of IP-DNA was determined by RT-qPCR using primer pairs amplifying a region around the NRE site in the IP-10 or IL-8 promoter (Table 2). To demonstrate the site specificity of the assay, a primer pair amplifying an irrelevant site around the 3′-UTR was also used. IgG ...
RIP-qPCR was performed using anti-YTHDF2 to confirm the interaction between c-Rel mRNA and YTHDF2 in all three PTC cell lines. Notably, YTHDF2 could also bind to the mRNA of RelA in PTC cells (Figure 5C). Given that YTHDF2 promotes mRNA degradation by selectively binding m6A-modified...
(G,H). Cells were harvested and viral DNA was extracted from cells and processed for qPCR analysis of viral DNA accumulation using HSV-1 DNA polymerase and UL26 primer for HSV-1 (I) and CMV (J), respectively. Real-time PCR with β-actin primers were performed to serve as an internal...
IL-8 expression was measured by qPCR and ELISA. A and B, moDCs were stimulated overnight with human secreted mucin (2 μg/ml) and IL-8 expression measured by qPCR (A) and ELISA (B). n≥ 5 independent experiments; *, p < 0.05 assessed using unpaired Student’s t test. C, ELISA...
Lungs were perfused with agarose. One lung was inflated for formalin fixation and paraffin embedding; the other lung was used to isolate genomic DNA for qPCR (Supplementary Table 2). Genomic DNA was isolated from the livers. The livers, tumours, and lymph nodes were sectioned and stained with...
RT-qPCR was performed using Power SYBR Green mix (Applied Biosystems) with ABI Prism 7300 sequence detector (Applied Biosystems). Cyclophilin A transcript levels were used for normalization of each target gene. The custom primers were designed with IDT Primer-Quest online software and the ...