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. 2020 Mar 25;12(4):772. doi: 10.3390/cancers12040772

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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PAICS in CRC is regulated and targeted by miR-128. (A) HCT116^p53-wt was transfected with NT-pre-miR, miR-128, miR-141, or miR-146 followed by the immunoblot analysis of PAICS protein expression. The numerical values below the blots indicate the normalized ratio of the PAICS/β-Actin. (B) The precursor miR-128 or NT pre-miR was co-transfected with luciferase constructs of either PAICS-3’UTR wild-type or mutant. The transfection of HCT116^p53-wt cells was accomplished with either NT-pre-miR or miR-128 to assess (C) cell proliferation, (D) colony formation, and (E) spheroid formation; scale bar—1000 µm. (F) An immunoblot analysis to show the restoration of the PAICS levels after the ectopic expression of miR-128 followed by co-transfection of precursor-miR-128 and inhibitor of miR-128 in SW480^p53-mut cells. Transfections were performed to assess the (G) colony formation and (H) cell invasion capacity.

PAICS in CRC is regulated and targeted by miR-128. (A) HCT116^p53-wt was transfected with NT-pre-miR, miR-128, miR-141, or miR-146 followed by the immunoblot analysis of PAICS protein expression. The numerical values below the blots indicate the normalized ratio of the PAICS/β-Actin. (B) The precursor miR-128 or NT pre-miR was co-transfected with luciferase constructs of either PAICS-3’UTR wild-type or mutant. The transfection of HCT116^p53-wt cells was accomplished with either NT-pre-miR or miR-128 to assess (C) cell proliferation, (D) colony formation, and (E) spheroid formation; scale bar—1000 µm. (F) An immunoblot analysis to show the restoration of the PAICS levels after the ectopic expression of miR-128 followed by co-transfection of precursor-miR-128 and inhibitor of miR-128 in SW480^p53-mut cells. Transfections were performed to assess the (G) colony formation and (H) cell invasion capacity.