American Society of Hematology

Fig. 3.

$Fig. 3. Retroviral expression of v-ErbA or v-ErbB in primary ebls and the consequences for ferritin biosynthesis. Bone marrow-derived, primary ebls expressing either the v-ErbA or the v-ErbB oncoprotein (see Materials and Methods) were kept for 24 hours under self-renewing conditions in the presence of 50 μmol/L Des to achieve iron depletion or were incubated with 1 mg/mL Tf to induce full iron saturation. (A) Distribution of ferH mRNA as determined by sucrose gradient analysis (for details, see legend to Fig 1). (Insets) The ribosome-bound mRNA compartment (fractions 9-18) is shown with an extended ordinate to highlight the difference between v-ErbA and v-ErbB expressing cells. (Open symbols) Des; (solid symbols) Tf. (B) Ferritin protein levels as detected by Western blotting in self-renewing primary ebls (SCF), v-ErbA– or v-ErbB–expressing ebls (v-ErbA, v-ErbB), and AEV-transformed red blood cells (HD3E22) under iron scarcity (Des) or high iron abundance (Tf). In case of SCF-progenitors, low Tf (0.03 mg/mL) instead of Des was used to induce IRP-activity. Numbers below each pair of lanes represent the factors of iron-dependent upregulation of ferritin expression (fold induction).$

Retroviral expression of v-ErbA or v-ErbB in primary ebls and the consequences for ferritin biosynthesis. Bone marrow-derived, primary ebls expressing either the v-ErbA or the v-ErbB oncoprotein (see Materials and Methods) were kept for 24 hours under self-renewing conditions in the presence of 50 μmol/L Des to achieve iron depletion or were incubated with 1 mg/mL Tf to induce full iron saturation. (A) Distribution of ferH mRNA as determined by sucrose gradient analysis (for details, see legend to Fig 1). (Insets) The ribosome-bound mRNA compartment (fractions 9-18) is shown with an extended ordinate to highlight the difference between v-ErbA and v-ErbB expressing cells. (Open symbols) Des; (solid symbols) Tf. (B) Ferritin protein levels as detected by Western blotting in self-renewing primary ebls (SCF), v-ErbA– or v-ErbB–expressing ebls (v-ErbA, v-ErbB), and AEV-transformed red blood cells (HD3E22) under iron scarcity (Des) or high iron abundance (Tf). In case of SCF-progenitors, low Tf (0.03 mg/mL) instead of Des was used to induce IRP-activity. Numbers below each pair of lanes represent the factors of iron-dependent upregulation of ferritin expression (fold induction).

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