Figure 5.
Suppression of p-S6 inhibited DLBCL cell proliferation and sensitized chemotherapy. (A) Primary mice fibroblasts (FRCs) were treated with the indicated concentration of Rapa for 24 hours. Relative cell viability was determined by CCK-8 assay. (B) FRCs were treated with the indicated concentrations of Rapa for 24 hours. Cell lysates were collected and the expression of p-S6 was determined by western blotting. (C) FRCs were treated with 1 nM of Rapa for 24 hours, and were collected at 1 to 4 days after withdrawal of Rapa. The expression of p-S6 was determined by western blotting. (D) FRCs were treated with 1 nM of Rapa for 24 hours before RNA-seq sample preparation. GO enrichment for downregulated genes of “Rapa-treated FRC” vs FRC. The red-green color scale corresponds to the enrichment significance of the genes. Dot size is proportional to the number of genes annotated to the corresponding categories. P values are calculated by Fisher exact test. (E) A schematic diagram of FRCs treatment with Rapa and coculture with DLBCL cells. (F) FRC lysates were collected to determine the protein expression of p-S6 and α-SMA. (G) FRCs were treated with shS6 to knock down the level of p-S6. (H) FRCs were pretreated with Rapa to suppress the expression of p-S6 and then cocultured with DB or A20 cells. DB or A20 cell number was counted by Countstar. (I) FRCs were pretreated with shS6 to suppress the expression of p-S6 and then cocultured with DB or A20 cells. DB or A20 cell number was counted by Countstar. (J) A schematic diagram of in vivo antitumor study using DB xenograft mice. (K) Tumors were weighted after dissection on day 9. (L) Photos of tumors were taken accompanied by the scale. DVC, DOX+VCR+CTX; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Rapa, rapamycin.

Suppression of p-S6 inhibited DLBCL cell proliferation and sensitized chemotherapy. (A) Primary mice fibroblasts (FRCs) were treated with the indicated concentration of Rapa for 24 hours. Relative cell viability was determined by CCK-8 assay. (B) FRCs were treated with the indicated concentrations of Rapa for 24 hours. Cell lysates were collected and the expression of p-S6 was determined by western blotting. (C) FRCs were treated with 1 nM of Rapa for 24 hours, and were collected at 1 to 4 days after withdrawal of Rapa. The expression of p-S6 was determined by western blotting. (D) FRCs were treated with 1 nM of Rapa for 24 hours before RNA-seq sample preparation. GO enrichment for downregulated genes of “Rapa-treated FRC” vs FRC. The red-green color scale corresponds to the enrichment significance of the genes. Dot size is proportional to the number of genes annotated to the corresponding categories. P values are calculated by Fisher exact test. (E) A schematic diagram of FRCs treatment with Rapa and coculture with DLBCL cells. (F) FRC lysates were collected to determine the protein expression of p-S6 and α-SMA. (G) FRCs were treated with shS6 to knock down the level of p-S6. (H) FRCs were pretreated with Rapa to suppress the expression of p-S6 and then cocultured with DB or A20 cells. DB or A20 cell number was counted by Countstar. (I) FRCs were pretreated with shS6 to suppress the expression of p-S6 and then cocultured with DB or A20 cells. DB or A20 cell number was counted by Countstar. (J) A schematic diagram of in vivo antitumor study using DB xenograft mice. (K) Tumors were weighted after dissection on day 9. (L) Photos of tumors were taken accompanied by the scale. DVC, DOX+VCR+CTX; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Rapa, rapamycin.

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