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. 2018 Mar 10;19(3):796. doi: 10.3390/ijms19030796

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© 2018 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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Regulation of TXNIP expression and redox balance in HL-60 cells by 1,25(OH)₂D₃ is glucose-dependent. (A,B) Immunoblots showing TXNIP protein levels. (A) In the absence of glucose, expression of TXNIP in HL-60 cells is significantly reduced, and appears to be unaffected by 1,25(OH)₂D₃ (100 nM) treatment. “+” and “−” denote the presence or absence of the indicated molecule/treatment, respectively; (B) In the presence of a single high dose of the glucose uptake inhibitor phloretin (200 µM), added at different time intervals prior to the end of the initial treatment period (24 h), 1,25(OH)₂D₃ is found to be incapable of inducing TXNIP expression; (C) Changes in TXNIP mRNA levels after 24 and 96 h of treatment with 1,25(OH)₂D₃ in HL-60 cells, cultured in the presence or absence of glucose, reveal that regulation by 1,25(OH)₂D₃ is glucose-dependent. The dashed red line indicates baseline mRNA expression in DMSO-treated cells set to 1. Error bars ± SD; n = 2; (D) Unlike TXNIP, the 1,25(OH)₂D₃ target gene CYP24A1, is induced in response to 1,25(OH)₂D₃ treatment (96 h) independent of glucose availability. Error bars ± SD; n = 2; (E) In absence of glucose, intracellular ROS levels are largely unaltered upon treatment of HL-60 cells for 96 h with 1,25(OH)₂D₃. The dashed red line indicates basal ROS levels in DMSO-treated cells set to 1. Statistical significance was calculated using a two-tailed Student’s t-test, with p-values less than or equal to 0.05 and 0.01 depicted by * and **, respectively. Nonsignificant results are denoted as n.s. Error bars ± SD; n = 3.

Regulation of TXNIP expression and redox balance in HL-60 cells by 1,25(OH)₂D₃ is glucose-dependent. (A,B) Immunoblots showing TXNIP protein levels. (A) In the absence of glucose, expression of TXNIP in HL-60 cells is significantly reduced, and appears to be unaffected by 1,25(OH)₂D₃ (100 nM) treatment. “+” and “−” denote the presence or absence of the indicated molecule/treatment, respectively; (B) In the presence of a single high dose of the glucose uptake inhibitor phloretin (200 µM), added at different time intervals prior to the end of the initial treatment period (24 h), 1,25(OH)₂D₃ is found to be incapable of inducing TXNIP expression; (C) Changes in TXNIP mRNA levels after 24 and 96 h of treatment with 1,25(OH)₂D₃ in HL-60 cells, cultured in the presence or absence of glucose, reveal that regulation by 1,25(OH)₂D₃ is glucose-dependent. The dashed red line indicates baseline mRNA expression in DMSO-treated cells set to 1. Error bars ± SD; n = 2; (D) Unlike TXNIP, the 1,25(OH)₂D₃ target gene CYP24A1, is induced in response to 1,25(OH)₂D₃ treatment (96 h) independent of glucose availability. Error bars ± SD; n = 2; (E) In absence of glucose, intracellular ROS levels are largely unaltered upon treatment of HL-60 cells for 96 h with 1,25(OH)₂D₃. The dashed red line indicates basal ROS levels in DMSO-treated cells set to 1. Statistical significance was calculated using a two-tailed Student’s t-test, with p-values less than or equal to 0.05 and 0.01 depicted by * and **, respectively. Nonsignificant results are denoted as n.s. Error bars ± SD; n = 3.