American Society of Hematology

Figure 6

$Figure 6. Enhanced cytokine production in ATF3-null MCs. WT or ATF3-null MCs were cultured in IL-3/SCF for 5 weeks, starved of growth factor for 6 hours, washed, and then sensitized with anti-DNP IgE and stimulated with DNP–bovine serum albumin (100 ng/mL) for 0, 1, and 6 hours. (A) Cell pellets were used for RNA isolation and real-time reverse transcription–PCR was performed to determine cytokine expression. (B) Cytokine release was determined in cell-free supernatants by enzyme-linked immunoabsorbent assay. (C) Chromatin immunoprecipitation analysis of ATF3 to predict binding sites in the IL-4 and IL-6 promoters was performed as outlined in “Chromatin immunoprecipitation” with the use of rabbit anti-ATF3 antibody. DNA from input or immunoprecipitated (ATF3) fractions was measured by PCR amplification of specific promoter sequences. Results are expressed as mean ± SEM for 3 independent experiments. *P < .05 by comparison to WT.$

Enhanced cytokine production in ATF3-null MCs. WT or ATF3-null MCs were cultured in IL-3/SCF for 5 weeks, starved of growth factor for 6 hours, washed, and then sensitized with anti-DNP IgE and stimulated with DNP–bovine serum albumin (100 ng/mL) for 0, 1, and 6 hours. (A) Cell pellets were used for RNA isolation and real-time reverse transcription–PCR was performed to determine cytokine expression. (B) Cytokine release was determined in cell-free supernatants by enzyme-linked immunoabsorbent assay. (C) Chromatin immunoprecipitation analysis of ATF3 to predict binding sites in the IL-4 and IL-6 promoters was performed as outlined in “Chromatin immunoprecipitation” with the use of rabbit anti-ATF3 antibody. DNA from input or immunoprecipitated (ATF3) fractions was measured by PCR amplification of specific promoter sequences. Results are expressed as mean ± SEM for 3 independent experiments. *P < .05 by comparison to WT.

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