Fig. 4: PRDM1 expression in ribosome-inactivated CRC cells.
From: Mucosal ribosomal stress-induced PRDM1 promotes chemoresistance via stemness regulation
a Wild type or APCMin/+ mice (n = 3–5) were treated with 25 mg/kg RIS-1 for 72 h, after which the PRDM1 levels in the small intestine were measured using immunohistochemistry with hematoxylin staining (magnification ×100; scale bar(s), 50 μm). The arrows indicate higher levels of PRDM1 expression in the crypt than in the villi. Quantification of PRDM1 staining from IHC analysis (the lower graph) and relative area of adenoma in the gut. Results are shown as a plot with Tukey whiskers and different letters (a–c) over each box represent significant differences between groups (the lower left graphs, n = 12–20, P < 0.05). Asterisks represent a significant difference compared to each vehicle group (the lower right graph, n = 16–39, P < 0.05). b PRDM1 mRNA expression was measured in the intestinal cancer cells treated with ID90 of RIS-1 or RIS-2 for 2 h. Results are shown as mean values ± SD and asterisks represent a significant difference compared to each vehicle group (n = 3, P < 0.05). c HCT-8 cells were transfected with control (the negative control shRNA) or shPKR plasmid. PRDM1 protein was detected in the HCT-8 cells treated with RIS-1 for 6 h, RIS-2 for 24 h or RIS-3 for 6 h (different times indicate points for each maximal expression of PRDM1). Total cell lysates were subjected to western blot analysis. d HCT-8 cells were treated with ID90 of RIS for 8 h (RIS-1 or RIS-3) or for 2 h (RIS-2) and then added 5 μM actinomycin D for the indicated time to arrest cellular transcription. Each mRNA level was measured using RT real-time PCR. Results are shown as mean values ± SD and asterisks (∗) indicate significant differences from the group without RIS treatment at each time point (n = 3, ***P < 0.001).
