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. 2020 May 16;13(8):100790. doi: 10.1016/j.tranon.2020.100790

Figure 1.

Figure 1

Reduced total and phosphorylated IGF1R protein levels in differentiated DCs. (A) Human leukemic THP-1 cells were differentiated to immature DCs by treatment with 100 ng/ml IL-4 and 100 ng/ml GM-CSF for 5 days and then differentiated to DCs by treatment with 200 ng/ml IL-4, 100 ng/ml GM-CSF, 20 ng/ml TNFα, and 200 ng/ml Ionomycin for 2 days. THP1 scale bar = 50 μm, immature DCs scale bar = 500 μm, DCs scale bar = 500 μm. (B) DC marker expression via CD11b and CD141 in THP-1, ImDC, and mDC. (C) THP-1 and THP-1 cells were treated with 200 ng/ml IL-4, 100 ng/ml GM-CSF, 20 ng/ml TNFα, and 200 ng/ml Ionomycin for 72 hours and were incubated with IGF1 for 10 minutes before harvest, after which cell extracts were prepared. Proteins were separated through SDS-PAGE, followed by electrophoretic transfer and incubation with antibodies against tIGF1R, pIGF1R, tAKT, and pAKT. (D) Scanning densitometry analysis of tIGF1R (blue bars) and pIGF1R (red bars) levels in THP-1 and DCs. A value of 100% was given to the expression level of untreated THP-1 cells. Levels of tubulin were measured as a loading control. *P < .05. Bars represent SEM values. (E) Scanning densitometry analysis of pAKT levels in THP-1 and DCs. A value of 100% was given to the expression level of untreated THP-1 cells. Levels of tubulin were measured as a loading control.*P < .05. Bars represent SEM values. The graphs represent average of three independent experiments. (F) Expression of tIGF1R and pIGF1R with (green) or without (blue) IGF1 treatment in THP-1 cells.