Antilymphoma effects of HSPH1 silencing in human aggressive B-NHL models. (A) Western blot analyses of HSPH1, Bcl-6, c-Myc, and actin as internal protein-loading control in the indicated wild-type (WT), MOCK, and siHSPH1 (siHSP) human aggressive B-NHL cell lines. Western blot images were acquired using Microtek ArtixScan F1 and cropped to retain the relevant bands with Adobe Photoshop CS version 4 for Macintosh computer. (B) Tumor growth curve (top) and tumor-free survival (middle, bottom) of SCID mice injected subcutaneously with 10⁶ or 10⁴ MOCK, siHSP_1, and siHSP_2 cells (6/group) (i). Tumor volume was calculated as 0.5 × d1²  × d2 (d1, smaller diameter; d2, larger diameter). Statistically significant differences were calculated by using the 2-way ANOVA with Bonferroni posttest or the log-rank (Mantel-Cox) test, respectively (*P < .05, **P < .01, ***P < .001). (ii) Quantitative software analyses of HSPH1, Bcl-6, and c-Myc expression evaluated by immunohistochemistry in MOCK, siHSP_1, and siHSP_2 xenografts 16 days after injection of 10⁶ cells. Data are shown as mean ± standard of the mean of the ratio between the immunostained area and the total nuclear area, measured by ImageJ software analysis v.146 in 3 to 5 high-power microscopic fields for each case. Significant differences were calculated with the unpaired 2-tailed Student t test (**P < .01, ***P < .001, ****P < .0001).