Fig. 6. Knockdown of TCF-4 inhibits TCTP-induced glioma cell proliferation in vitro and in vivo. (A) TCF-4 was knocked down in U-251/TCTP and H-4/TCTP.

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Fig. 6. Knockdown of TCF-4 inhibits TCTP-induced glioma cell proliferation in vitro and in vivo. (A) TCF-4 was knocked down in U-251/TCTP and H-4/TCTP cells. U-251/TCTP cells and H-4/TCTP cells were transfected with control shRNA (Ctrl shRNA) or TCF-4 shRNA1. TCTP levels were detected through immunostaining with the TCTP antibody, and actin was used as a loading control. (B) In vitro growth of U-251/TCTP cells treated with nonsense control shRNA (Ctrl shRNA), or TCF-4 shRNA1 was measured using MTT assays. (C) In vitro growth of H-4/TCTP cells treated with nonsense control shRNA (Ctrl shRNA), or TCF-4 shRNA1 was measured using MTT assays. (D) In vitro growth of U-251/V cells treated with nonsense control shRNA (Ctrl shRNA) or TCF-4 shRNA1 was measured using MTT assays. (E) In vitro growth of H-4/V cells treated with nonsense control shRNA (Ctrl shRNA), or TCF-4 shRNA1 was measured using MTT assays. (F) Average tumor volume (left) and tumor TCF-4/Ki67 staining (5 weeks after implantation, right) in athymic nude mice subcutaneously inoculated with U-251/TCTP treated with nonsense control shRNA (Ctrl shRNA) or TCF-4 shRNA1. (G) Average tumor volume (left) and tumor TCF-4/Ki67 staining (5 weeks after implantation, right) in athymic nude mice subcutaneously inoculated with H-4/TCTP treated with nonsense control shRNA (Ctrl shRNA) or TCF-4 shRNA1. (H) Another TCF-4 shRNA called TCF-4 shRNA2 was used to knock down TCF-4. U-251/TCTP, and H-4/TCTP cells were transfected with control shRNA (Ctrl shRNA) or TCF-4 shRNA2. TCTP levels were detected through immunostaining with TCTP antibody, and actin was used as a loading control. (I) In vitro growth of U-251/TCTP cells treated with nonsense control shRNA (Ctrl shRNA), or TCF-4 shRNA2 was measured using MTT assays. (J) In vitro growth of H-4/TCTP cells treated with nonsense control shRNA (Ctrl shRNA), or TCF-4 shRNA2 was measured using MTT assays. (K) In vitro growth of U-251/V cells treated with nonsense control shRNA (Ctrl shRNA) or TCF-4 shRNA2 was measured using MTT assays. (L) In vitro growth of H-4/V cells treated with nonsense control shRNA (Ctrl shRNA), or TCF-4 shRNA2 was measured using MTT assays. (M) Average tumor volume (left) and tumor TCF-4/Ki67 staining (5 weeks after implantation, right) in athymic nude mice subcutaneously inoculated with U-251/TCTP treated with nonsense control shRNA (Ctrl shRNA) or TCF-4 shRNA2. (N) Average tumor volume (left) and tumor TCF-4/Ki67 staining (5 weeks after implantation, right) in athymic nude mice subcutaneously inoculated with H-4/TCTP treated with nonsense control shRNA (Ctrl shRNA) or TCF-4 shRNA2. For A–E and HL), the results represent at least 3 separate experiments. For F, G, M, and N, n = 10 mice/group. Error bar: ±S.D. From: TCTP promotes glioma cell proliferation in vitro and in vivo via enhanced β-catenin/TCF-4 transcription Neuro Oncol. 2013;16(2):217-227. doi:10.1093/neuonc/not194 Neuro Oncol | © The Author(s) 2013. Published by Oxford University Press on behalf of the Society for Neuro-Oncology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com. 1