Fig 1: HOXB2 activates the expression of CCT6A in colon cancer cells. (A and B) The binding sites between HOXB2 and CCT6A. The expression of HOXB2 in colon cancer cells was determined with (C) western blotting and (D) RT-qPCR. The expression of HOXB2 in colon cancer cells of the overexpression group was determined with (E) RT-qPCR and (F) western blotting. (G) A luciferase reporter assay was performed to detect the relationship between CCT6A and HOXB2. (H) Chromatin immunoprecipitation assays were performed to demonstrate the interaction between CCT6A and HOXB2. The expression of CCT6A in colon cancer cells was detected with (I) RT-qPCR and (J) western blotting. ***P<0.001. CCT6A, chaperonin containing TCP1 subunit 6A; RT-qPCR, reverse transcription-quantitative PCR; Ov, overexpressing; NC, negative control; WT, wild-type; MUT, mutant; shRNA, short hairpin RNA.
Fig 2: The presence of SE–TF regulatory network consisting of SMAD3, ETS1, and HOXB2 in T24 cells. (a) H3K27ac intensity of SMAD3 in eight common malignant tumor cell lines analyzed by ChIP-PCR. (b) Expression of core TFs of bladder cancer (SMAD3, ETS1, and HOXB2) in T24 cells determined by RT-qPCR. (c) Silencing and overexpression efficiency of SMAD3, ETS1, and HOXB2 in T24 cells determined by western blot analysis. (d) Overexpression efficiency of SMAD3, ETS1, and HOXB2 in T24 cells determined by RT-qPCR. (e) Expression of other two TFs in T24 cells following silencing of any bladder cancer TFs determined by western blot analysis. (f) Expression of other two TFs in T24 cells following overexpression of any bladder cancer TFs determined by RT-qPCR. (g) Expression of other TFs in T24 cells following intervention with two types of bladder cancer TFs determined by western blot analysis. (h) Expression of other TFs in T24 cells following intervention with two types of bladder cancer TFs determined by RT-qPCR. (i) The binding of each TF to the promoter regions of the other two TFs analyzed by ChIP-PCR. *p < 0.05. The experiment was conducted three times independently.
Fig 3: The SE–TF regulatory network consisting of SMAD3, ETS1, and HOXB2 promotes the malignant phenotype of bladder cancer cells. (a) Migration, invasion, and proliferation of T24 cells following overexpression or silencing of SMAD3, ETS1, and HOXB2 measured by Transwell and CCK-8 assays. (b) Migration, invasion, and proliferation of T24 cells following transfection with sh-SMAD3, sh-SMAD3 + pcDNA3.1-ETS1, sh-SMAD3 + pcDNA3.1-HOXB2, or sh-SMAD3 + pcDNA3.1-ETS1 + pcDNA3.1-HOXB2 measured by Transwell and CCK-8 assays. *p < 0.05. The experiment was conducted three times independently.
Fig 4: Overexpression of HOXB2 rescues the proliferation of colon cancer cells. (A) Cell Counting Kit-8 was performed to detect the viability of colon cancer cells. (B) The proliferation of colon cancer cells was determined with the colony formation assay. (C) Immunofluorescence was performed to detect the expression of Ki-67 in colon cancer cells. *P<0.05, **P<0.01 and ***P<0.001. ##P<0.01 vs. shRNA-CCT6A + Ov-NC. CCT6A, chaperonin containing TCP1 subunit 6A; shRNA, short hairpin RNA; Ov, overexpressing; NC, negative control.
Fig 5: Overexpression of HOXB2 rescues the invasion of colon cancer cells. (A) Wound healing and (B) Transwell assays were performed to determine the migration and invasion of colon cancer cells. (C) Western blotting was used for the detection of the expression of epithelial-mesenchymal transition-related proteins in colon cancer cells. **P<0.01 and ***P<0.001. CCT6A, chaperonin containing TCP1 subunit 6A; shRNA, short hairpin RNA; Ov, overexpressing; NC, negative control.
Supplier Page from Abcam for Anti-HOXB2 antibody