American Society of Hematology

Figure 4.

$DKK1 activates NF-KB signaling pathway by CKAP4 receptor in multiple myeloma. (A) Fold changes in activations of 17 signaling pathways treated with PBS or rhDKK1 (100 ng/mL) for 24 hours in MM.1S cells. (B) Phosphorylation levels of p65 and STAT3, total IκB-α in MM.1S and OPM-2 cells treated with increasing dosage of rhDKK1 (0-100 ng/mL) for 1 hour. (C) Immunofluorescence staining to detect the nuclear localization of p65 in MM.1S and OPM-2 cells treated with PBS or rhDKK1 (100 ng/mL) for 12 hours. Scale bar, 5 μm. (D) Cytoplasmic and nuclear fraction of p65 in MM.1S and OPM-2 cells treated with PBS or rhDKK1 (100 ng/mL) for 12 hours. (E) qPCR detecting downstream target genes of NF-κB signaling under the stimulation of rhDKK1 (100 ng/mL) for 12 hours. (F) Levels of p-p65 and IκB-α protein in MM.1S and OPM-2 cells treated with full-length (FL) rhDKK1 or CRD1 domain depletion DKK1 (ΔCRD1) for 12 hours. (G) Levels of p-p65 and IκB-α protein in MM.1S and OPM-2 cells with CKAP4 receptor knockdown and treated with rhDKK1 (100 ng/mL) for 12 hours. (H) Immunohistochemistry staining of DKK1 and p65 proteins in the sequential slides of bone marrow biopsies from 2 patients and healthy donor. Scale bar, 20 μm. (I) Correlation coefficient between positive rate of p65 and DKK1 intensity in bone marrow tissues from MM patients (n = 12).$

DKK1 activates NF-KB signaling pathway by CKAP4 receptor in multiple myeloma. (A) Fold changes in activations of 17 signaling pathways treated with PBS or rhDKK1 (100 ng/mL) for 24 hours in MM.1S cells. (B) Phosphorylation levels of p65 and STAT3, total IκB-α in MM.1S and OPM-2 cells treated with increasing dosage of rhDKK1 (0-100 ng/mL) for 1 hour. (C) Immunofluorescence staining to detect the nuclear localization of p65 in MM.1S and OPM-2 cells treated with PBS or rhDKK1 (100 ng/mL) for 12 hours. Scale bar, 5 μm. (D) Cytoplasmic and nuclear fraction of p65 in MM.1S and OPM-2 cells treated with PBS or rhDKK1 (100 ng/mL) for 12 hours. (E) qPCR detecting downstream target genes of NF-κB signaling under the stimulation of rhDKK1 (100 ng/mL) for 12 hours. (F) Levels of p-p65 and IκB-α protein in MM.1S and OPM-2 cells treated with full-length (FL) rhDKK1 or CRD1 domain depletion DKK1 (ΔCRD1) for 12 hours. (G) Levels of p-p65 and IκB-α protein in MM.1S and OPM-2 cells with CKAP4 receptor knockdown and treated with rhDKK1 (100 ng/mL) for 12 hours. (H) Immunohistochemistry staining of DKK1 and p65 proteins in the sequential slides of bone marrow biopsies from 2 patients and healthy donor. Scale bar, 20 μm. (I) Correlation coefficient between positive rate of p65 and DKK1 intensity in bone marrow tissues from MM patients (n = 12).

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