Fig 1: CIRP leads to increased inflammatory process in the PBMCs by rs11170510 and rs58123204. (A) THP-1 cells were transfected with the Sp1 plasmid for 24 (h) Immunofluorescence microscopy analysis of CIRP. Merged signals of CIRP (Alexa Fluor 488-conjugated goat anti-rabbit IgG, green) and DAPI (nuclei, blue). scale bar: 10 um. (B) Quantification of translocation in then nucleus to cytosol foci per cell. (C) CIRP levels were measured in serum with major allele or minor allele of the rs11170510 and rs58123204 (n = 10, major homo AA; n = 10, minor homo GG) using CIRP ELISA kit. (D) PBMCs were analyzed using the quantitative PCR of Tnf-a, Il6, Il-1ß, and Il8 mRNA levels. (E) C-reactive protein levels were measured in serum containing major A allele or minor G allele of rs11170510 and rs58123204 (n = 10, major homo AA; n = 10, minor homo GG). Experiments were repeated at least three times. Graphs are indicative of the mean ± SD. **p < 0.01 and ***p < 0.001 (two-tailed unpaired Mann–Whitney).
Fig 2: SP1 regulates the transcriptional activation of CIRP by rs11170510 and rs58123204. (A) A strong transcription factor cluster appeared in the Cirp transcriptional promoter region. The SP1 transcription factor binding site on the Cirp promoter was located in the region chr19:1269276-1269290. (B) HEK293 cells were transfected with a Cirp luciferase reporter construct in the presence or absence of plasmid Sp1 (0.1, 0.5, and 1µg). Cells were subjected to luciferase assays. (C) HEK293 cells were transfected plasmid Sp1 (1 µg) and analyzed using quantitative PCR for Cirp mRNA expression. (D) PBMCs with major A allele or minor G allele of rs11170510 and rs58123204 were subjected to quantitative PCR for Cirp mRNA using its primer (n = 10, major homo AA; n = 10, minor homo GG). Experiments were repeated at least three times. Graphs are indicative of the mean ± SD. *p < 0.05, **p < 0.01 and ***p < 0.001 (two-tailed unpaired Mann–Whitney).
Supplier Page from CUSABIO Technology LLC for Human Cold-inducible RNA-binding protein(CIRBP) ELISA Kit