Figure 7
Figure 7. RI-BPI inhibits BCL6 transcriptional repression and induces DLBCL apoptosis in vivo. (A) Representative images from SU-DHL4 and SU-DHL6 mice tumors after being treated with control peptide (first column), RI-BPI 150 μg/day (second column), or RI-BPI 500 μg/day (third column), and assayed for apoptosis by TUNEL. The plot on the far right represents the apoptotic index (apoptotic cells over total cells) with the percentage of apoptotic cells in the y-axis for pooled control peptide doses (■), RI-BPI 150 μg/day (□), and RI-BPI 500 μg/day () in the x-axis. (B) The same tumors as in panel A were assayed for proliferation by proliferating cell nuclear antigen immunostaining. Nuclei were classified as negative, low-intensity positive, and high-intensity positive as shown in the inset (digital zoom) of the SUDHL4 control tumor, by green, yellow, or red circles, respectively. The plot on the far right represents the proliferation index (using the same color coding for negative and positive cells) with the percentage of proliferating and nonproliferating cells in the y-axis for pooled control peptide samples (first stacking column), RI-BPI 150 μg/day (second stacking column), and RI-BPI 500 μg/day (third stacking column). (C) The same tumors were examined for the presence of mitotic cells. The plot on the far right represents the mitotic index (mitotic cells over total cells) with the percentage of mitotic cells in the y-axis in pooled control peptide samples (■), RI-BPI 150 μg/day (□), and RI-BPI 500 μg/day (). (A-C) Slides were mounted with permanent mounting medium (Vectamount; Vector Laboratories). Slides were viewed with a light microscope (AxioSkop 2; Carl Zeiss) using a Plan-neofluar lens at a 10×/0.50 air objective, a 25×/0.80 oil objective, a 40×/0.90 oil objective, and a 100×/1.30 oil objective. Images were acquired using a color camera (AxioCam; Carl Zeiss), processed using Axiovision software (Carl Zeiss), and scored using ImageJ software (National Institutes of Health, Bethesda, MD). (D) The mRNA abundance of TP53 and ATR was determined in the same tumors by quantitative RT-PCR. The y-axis represents the normalized amount of mRNA in arbitrary units as measured by the relative standard curve method.

RI-BPI inhibits BCL6 transcriptional repression and induces DLBCL apoptosis in vivo. (A) Representative images from SU-DHL4 and SU-DHL6 mice tumors after being treated with control peptide (first column), RI-BPI 150 μg/day (second column), or RI-BPI 500 μg/day (third column), and assayed for apoptosis by TUNEL. The plot on the far right represents the apoptotic index (apoptotic cells over total cells) with the percentage of apoptotic cells in the y-axis for pooled control peptide doses (■), RI-BPI 150 μg/day (□), and RI-BPI 500 μg/day () in the x-axis. (B) The same tumors as in panel A were assayed for proliferation by proliferating cell nuclear antigen immunostaining. Nuclei were classified as negative, low-intensity positive, and high-intensity positive as shown in the inset (digital zoom) of the SUDHL4 control tumor, by green, yellow, or red circles, respectively. The plot on the far right represents the proliferation index (using the same color coding for negative and positive cells) with the percentage of proliferating and nonproliferating cells in the y-axis for pooled control peptide samples (first stacking column), RI-BPI 150 μg/day (second stacking column), and RI-BPI 500 μg/day (third stacking column). (C) The same tumors were examined for the presence of mitotic cells. The plot on the far right represents the mitotic index (mitotic cells over total cells) with the percentage of mitotic cells in the y-axis in pooled control peptide samples (■), RI-BPI 150 μg/day (□), and RI-BPI 500 μg/day (). (A-C) Slides were mounted with permanent mounting medium (Vectamount; Vector Laboratories). Slides were viewed with a light microscope (AxioSkop 2; Carl Zeiss) using a Plan-neofluar lens at a 10×/0.50 air objective, a 25×/0.80 oil objective, a 40×/0.90 oil objective, and a 100×/1.30 oil objective. Images were acquired using a color camera (AxioCam; Carl Zeiss), processed using Axiovision software (Carl Zeiss), and scored using ImageJ software (National Institutes of Health, Bethesda, MD). (D) The mRNA abundance of TP53 and ATR was determined in the same tumors by quantitative RT-PCR. The y-axis represents the normalized amount of mRNA in arbitrary units as measured by the relative standard curve method.

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