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. 2024 Feb 12;212(7):1207–1220. doi: 10.4049/jimmunol.2300651

FIGURE 7.

FIGURE 7.

LcCRFB1 and LcCRFB5 receptor blockade impairs LcIFNd signal transduction. (A) a, Left, SDS-PAGE gel showing purified rLcCRFB1-Ex. Right, Detection of endogenous full-length LcCRFB1 from PKL cells (lane 1) and spleen cells (lane 2) and rLcCRFB1-Ex (lane 3) and control (lane 4). The theoretical molecular masses of full-length LcCRFB1 and LcCRFB1-Ex are 59.62 and 22.76 kDa, respectively. b, Verification of anti–LcCRFB1-Ex pAb specificity by immunofluorescence labeling in PKL and HEK-293T cells. The pCMV-HA plasmid was overexpressed in HEK-293T cells as control, and pCMV-HA-LcCRFB1 containing full-length LcCRFB1 cDNA was overexpressed in HEK-293T cells as the experimental group. (B) JAK1, IRF9, STAT1, and STAT2 and (C) MxA, PKR, and viperin expression levels upon receptor blockade with anti–LcCRFB1-Ex, anti–LcCRFB5-Ex, or both pAbs in LcIFNd-stimulated PKLs. (D) STAT1 and STAT2 phosphorylation were inhibited by anti–LcCRFB1-Ex, anti–LcCRFB5-Ex, or both Abs in LcIFNd-stimulated PKLs. Bars represent the means of three independent experiments ± SD. All experiments were performed in triplicate. Different lowercase letters indicate significant differences. **p < 0.01.