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Fig. 2 | Journal of Hematology & Oncology

Fig. 2

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

Fig. 2

Gfi1 mediates cell growth and viability of myeloma cells. mRNA Gfi1 levels measured by qPCR showing its knockdown (KD) as compared with scrambled (Scr) control in H929 cells using two different lentiviral shRNA (#1 and #2) (a). p53 target genes (BAX, NOXA, PUMA) mRNA levels were measured by qPCR in Gfi1-KD and Scr control H929 cells. The bar graph represents fold versus Scr levels (b). Gfi1, BAX, NOXA, and PUMA protein levels were analyzed by WB using GAPDH as loading control (left panel). The graph (right panel) represents densitometric levels of the protein versus loading control in three independent experiments (c). H929 cells were transduced with Gfi1-shRNA (#1 and #2) and Scr-shRNA, selected with puromycin for 48 h and maintained in complete media for another 24 h. The cell number was counted by hemocytometer (time zero) and after another 24 and 72 h. The bar graph represents percent cell number vs time zero (d). Gfi1, BAX, total and cleaved caspase 3 protein levels were analyzed by WB using α-tubulin as loading control (e) and DNA fragmentation was measured by Apo-BRDU assay and flow cytometry in H929 cells transduced with Gfi1-shRNA (#1) and Scr-shRNA (f)

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