Figure 6
Figure 6. Ly49D, IL-12, and IL-18 stimulation elevates IL-15 transcription. (A) Total B cells from Ii−/− (immature) mice were incubated in the presence or absence of IL-15 (30 ng/mL) for various periods of time, as indicated, and RT-PCR was carried out using primers for IFN-γ, Ly49D, and HPRT. (B) Total B cells from Ii−/− mice (immature) incubated in the presence or absence of anti-Ly49D antibody or an isotype control antibody RT-PCR was carried out using primers for IL-15, IFN-γ, and HPRT. (C,D) Total B cells from Ii−/− mice (immature) incubated in the presence or absence of anti-Ly49D antibody or an isotype control antibody. Quantitative real-time PCR was carried out using primers for IL-15 (C) or for IFN-γ (D). (E) Total B cells from Ii−/− mice (immature) were incubated in the presence or absence of IL-12 (20 ng/mL) and IL-18 (200 ng/mL). RT-PCR was carried out using primers for IL-15, IFN-γ, and HPRT. (F,G) Total B cells from Ii−/− mice (immature) were incubated in the presence or absence of IL-12 (20 ng/mL) and IL-18 (200 ng/mL), and quantitative real-time PCR was carried out using primers for IL-15 (F) or for IFN-γ (G). (H) Total B cells from Ii−/− (immature) mice were incubated overnight in the presence or absence of anti-Ly49D antibody, or IL-12 (20 ng/mL) and IL-18 (200 ng/mL). IL-15 levels in the conditioned medium of the cells was analyzed by ELISA. The results presented are representative of 3 separate experiments. Standard error was calculated. (I) Total B cells from control and IL-15−/− mice were incubated in the presence of absence of IL-12 (20 ng/mL) and IL-18 (200 ng/mL), and RT-PCR was carried out using primers for IFN-γ and HPRT. The results are representative of 3 separate experiments. (J) Total B cells from control and IL-15−/− mice were incubated in the presence or absence of Ly49D, and quantitative real-time PCR was carried out using primers for IFN-γ and HPRT. The results are representative of 3 separate experiments. (K) Purified immature B cells (CD21−, B220+) from IL-15−/− mice were incubated in the presence of anti-Ly49D for 3 hours. After incubation, their actin polymerization was analyzed as described in “Cytoskeleton rearrangement.” Error bars represent SD.

Ly49D, IL-12, and IL-18 stimulation elevates IL-15 transcription. (A) Total B cells from Ii−/− (immature) mice were incubated in the presence or absence of IL-15 (30 ng/mL) for various periods of time, as indicated, and RT-PCR was carried out using primers for IFN-γ, Ly49D, and HPRT. (B) Total B cells from Ii−/− mice (immature) incubated in the presence or absence of anti-Ly49D antibody or an isotype control antibody RT-PCR was carried out using primers for IL-15, IFN-γ, and HPRT. (C,D) Total B cells from Ii−/− mice (immature) incubated in the presence or absence of anti-Ly49D antibody or an isotype control antibody. Quantitative real-time PCR was carried out using primers for IL-15 (C) or for IFN-γ (D). (E) Total B cells from Ii−/− mice (immature) were incubated in the presence or absence of IL-12 (20 ng/mL) and IL-18 (200 ng/mL). RT-PCR was carried out using primers for IL-15, IFN-γ, and HPRT. (F,G) Total B cells from Ii−/− mice (immature) were incubated in the presence or absence of IL-12 (20 ng/mL) and IL-18 (200 ng/mL), and quantitative real-time PCR was carried out using primers for IL-15 (F) or for IFN-γ (G). (H) Total B cells from Ii−/− (immature) mice were incubated overnight in the presence or absence of anti-Ly49D antibody, or IL-12 (20 ng/mL) and IL-18 (200 ng/mL). IL-15 levels in the conditioned medium of the cells was analyzed by ELISA. The results presented are representative of 3 separate experiments. Standard error was calculated. (I) Total B cells from control and IL-15−/− mice were incubated in the presence of absence of IL-12 (20 ng/mL) and IL-18 (200 ng/mL), and RT-PCR was carried out using primers for IFN-γ and HPRT. The results are representative of 3 separate experiments. (J) Total B cells from control and IL-15−/− mice were incubated in the presence or absence of Ly49D, and quantitative real-time PCR was carried out using primers for IFN-γ and HPRT. The results are representative of 3 separate experiments. (K) Purified immature B cells (CD21−, B220+) from IL-15−/− mice were incubated in the presence of anti-Ly49D for 3 hours. After incubation, their actin polymerization was analyzed as described in “Cytoskeleton rearrangement.” Error bars represent SD.

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