Human CCL22/MDC Antibody

Detection of Human CCL22/MDC by Western Blot

MiR‐23a inhibited Tregs migration by directly targeting CCL22. A, CCL22 3′UTR contains a putative miR‐23a‐binding site. B, MiR‐23a mimics significantly reduced the luciferase activity tagged with wide type of CCL22 3′UTR. Mutagenesis in the miR‐23a‐binding site abolished the inhibitory actions. C, Transfection of miR‐23a mimics efficiently increased miR‐23a level in both cell lines. D, The effects of miR‐23a overexpression on p‐p65, p65, and CCL22 protein levels in HepG2 and HepG2.2.15 cells were assessed by western blotting. E, The gray scale analysis of p‐p65, p65, CCL22 after miR‐23a overexpression. MiR‐23a mimics transfection did not alter the expression of p65 and its phosphorylation but CCL22 was significantly reduced by miR‐23a mimics. F, MiR‐23a mimics significantly reduced CCL22 mRNA level. G, Overexpression of miR‐23a was sufficient to attenuate Tregs transmigration in both cell lines. Error bars represented mean ± SD. *P < .05, **P < .01, and ***P < .001 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31769216), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CCL22/MDC by Western Blot

Lower miR‐23a expression was correlated with higher level of CCL22 expression and intratumoral Treg recruitment in HBV‐positive HCC. A, HBV infection was associated with patient survival rate of HCC. Patients carrying HBV (n = 30) had significantly poorer prognosis than HBV‐ patients (n = 30). B, HBV‐ tissues (n = 30) and HBV+ tissues (n = 30) expressed significantly lower level of miR‐23a. C, HBV− tissues and HBV+ tissues expressed significantly higher mRNA level of CCL22. D, HBV‐ tissues and HBV+ tissues expressed significantly higher mRNA level of Foxp3. E, The protein levels of CCL22, Foxp3, p‐p65, and p65 in normal control, HBV‐ and HBV+ tumor tissues were evaluated by western blotting. F, The gray scale analysis of CCL22, Foxp3, p‐p65, and p65 in normal control, HBV− and HBV+ tumor tissues. In ascending order: normal < HBV− < HBV+. G, Foxp3 signals (red) were colocalized with CD4 (green) signals in tissues. The ratios of Foxp3+CD4+ cells were gradually increased from normal, HBV− HCC to HBV+ HCC tissues. H. MiR‐23a level was inversely correlated with CCL22 expression in HCC tissues, but not in normal control. I. MiR‐23a level was inversely correlated with Foxp3 expression in HCC tissues, but not in normal control. Error bars represented mean ± SD. **P < .01 and *P < .05. HBV+, HCC tissues with HBV infection. HBV−, HCC tissues without HBV infection. Normal, normal liver samples Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31769216), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CCL22/MDC by Western Blot

MiR‐23a inhibitor reversed p65 inhibition effects on CCL22 and Tregs recruitment. A, MiR‐23a inhibitor reversed p65 inhibition effects on miR‐23a level. B, The effects of miR‐23a inhibition and p65 inhibitor treatment on p‐p65, p65, and CCL22 protein levels in HepG2 and HepG2.2.15 cells were assessed by western blotting. C, The gray scale analysis of p‐p65, p65, CCL22. MiR‐23a inhibitor reversed p65 inhibition effects on CCL22 protein level but had no effects on p‐p65 and p65. D, MiR‐23a inhibitor reversed p65 inhibition effects on CCL22 mRNA level. E, MiR‐23a inhibitor reversed p65 inhibition effects on Tregs recruitment. F, CCL22 neutralizing antibody reversed miR‐23a inhibitor‐mediated Tregs migration. Error bars represented mean ± SD. *P < .05, **P < .01, and ***P < .001 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31769216), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CCL22/MDC by Western Blot

p65 promoted Tregs recruitment via directly modulating miR‐23a. A, A p65‐binding site could be found in miR‐23a promoter region. B, Overexpression of p65 significantly reduced the luciferase activity driven by miR‐23a promoter. Mutagenesis in the p65‐binding site released such repression. C, ChIP assay confirmed that p65 could bind to the promoter region of miR‐23a. D, p65 inhibition upregulated miR‐23a level. E, p65 inhibitor significantly reduced the p‐p65, p65, and CCL22 protein levels in HepG2 and HepG2.2.15 cells. F, The gray scale analysis of p‐p65, p65, CCL22 after p65 inhibitor treatment. G, p65 inhibition dampened CCL22 mRNA level. H, CD4+CD25+ Foxp3+ human Tregs were identified. I, HBV+ HepG2.2.15 cells caused more Tregs to migrate through transwell membrane than HBV− HpeG2 cells. Pretreatment with p65 inhibitor attenuated Tregs transmigration in both cell lines. Error bars represented mean ± SD. *P < .05, **P < .01, and ***P < .001 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31769216), licensed under a CC-BY license. Not internally tested by R&D Systems.