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Fig. 5 | Journal of Hematology & Oncology

Fig. 5

From: Growth factor independence 1 expression in myeloma cells enhances their growth, survival, and osteoclastogenesis

Fig. 5

Microenvironmental factors increase Gfi1 levels in MM cells. MM cells (H929) were direct co-cultured for 24 h with marrow stromal cells (BMSCs) obtained from normal donors. MM cells were harvested separate, and Gfi1 mRNA levels were measured by qPCR and compared with levels in MM cells alone (N = 4) (a, left panel). H929 cell lysates were analyzed by WB after 24 h direct co-culture with patient BMSC (MM-BMSC) using anti-Gfi1 and Mcl-1 antibodies and β actin as a loading control (a, right panel). MM cells with different p53 status: wt (H929 and MM.1S); haploinsufficient (JJN3) and null (KMS-11) were treated for 4 h with IL6, TNFα, and S1P at the indicated concentrations. The bar graph represents Gfi1 mRNA levels detected by real-time PCR and expressed as fold change versus untreated cells (b). H929 and MM.1S cells were treated with and without IL6 (5 ng/ml) for 4 h. Gfi1 and Mcl-1 protein levels in the cell lysates were analyzed by WB using β actin as a loading control (c). Gfi1-KD and Scr control H929 cells were direct co-cultured for 24 h with patient BMSC (MM-BMSC) or treated for 4 h with IL-6 (5 ng/ml). Gfi1 and Mcl-1 protein levels in the cell lysates were analyzed by WB using β actin as a loading control (d)

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