American Society of Hematology

Figure 6

$Figure 6. The small GTPase Rac1 is involved in simvastatin-enhanced IL-1β secretion. (A) THP-1 cells were serum-starved overnight and subsequently cultured in the presence or absence of simvastatin (10 μM) for 48 hours. GTPase pulldown fractions and cell lysates were assayed for Rac1 content. (B) Mevalonate (1 mM) was added just before simvastatin. Actin content served as control for equal protein loading. Vertical lines have been inserted to indicate a repositioned gel lane, within the same gel. Shown are representative blots of at least 3 independent experiments. (C) THP-1 cells were cultured for 24 hours in the presence or absence of simvastatin (10 μM) and stimulated for an additional 4 hours with 200 ng/mL LPS. Rac1 inhibitor was added either 1 hour before simvastatin or 1 hour before LPS treatment. Release of IL-1β was determined by ELISA; data are mean plus or minus SEM (n = 5; *P < .01). (D) THP-1 cells were serum-starved overnight and subsequently cultured in the presence or absence of simvastatin (10 μM) for 48 hours. Rac1 inhibitor was added 1 hour before simvastatin incubation. After the incubation, cells were lysed by adding lysis buffer and assayed for phospho-PKB and total PKB content. Actin content served as control for equal protein loading. Shown are representative blots of at least 3 independent experiments.$

The small GTPase Rac1 is involved in simvastatin-enhanced IL-1β secretion. (A) THP-1 cells were serum-starved overnight and subsequently cultured in the presence or absence of simvastatin (10 μM) for 48 hours. GTPase pulldown fractions and cell lysates were assayed for Rac1 content. (B) Mevalonate (1 mM) was added just before simvastatin. Actin content served as control for equal protein loading. Vertical lines have been inserted to indicate a repositioned gel lane, within the same gel. Shown are representative blots of at least 3 independent experiments. (C) THP-1 cells were cultured for 24 hours in the presence or absence of simvastatin (10 μM) and stimulated for an additional 4 hours with 200 ng/mL LPS. Rac1 inhibitor was added either 1 hour before simvastatin or 1 hour before LPS treatment. Release of IL-1β was determined by ELISA; data are mean plus or minus SEM (n = 5; *P < .01). (D) THP-1 cells were serum-starved overnight and subsequently cultured in the presence or absence of simvastatin (10 μM) for 48 hours. Rac1 inhibitor was added 1 hour before simvastatin incubation. After the incubation, cells were lysed by adding lysis buffer and assayed for phospho-PKB and total PKB content. Actin content served as control for equal protein loading. Shown are representative blots of at least 3 independent experiments.

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