1. Retinoids Bias Integrin Expression and Function in Cutaneous T-Cell Lymphoma 
Lei Wang, Sebastian S. DeMarco, JianMing Chen, Charles M. Phillips, Lance C. Bridges 
Journal of Investigative Dermatology 
Volume 135, Issue 8, Pages (August 2015)
DOI: /jid
Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

2. Figure 1
Retinoids selectively induce integrin expression in cutaneous T-cell lymphoma (CTCL) cell lines. Indicated T-cell lines were cultured for 72 hours in the presence of 100 nM all-trans-retinoic acid (ATRA; gray), 1 μM Bexarotene (black), or equimolar concentration of the vehicle DMSO (white). Cells were harvested and assessed for integrin β-subunit expression.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
Retinoids augment integrin β7 function in cutaneous T-cell lymphoma (CTCL) cell lines. (a) T-cell lines were incubated for 72 hours with DMSO or 100 nM all-trans-retinoic acid (ATRA). Adhesion to 0.75 μg ml−1 of immobilized mucosal addressin cell adhesion molecule (MAdCAM)-1-Fc was determined. Ordinate values (average A405) represent the relative extent of adhesion. Assay buffer included MnCl2 to uniformly activate integrin receptors. (b) Adhesion assays were repeated as in a with cells exposed to 1 μM Bexarotene. (c) CTCL cells cultured for 72 hours in the presence of 100 nM ATRA (gray), 1 μM Bexarotene (black), or vehicle (white) were added to wells coated with 0.5 μg ml−1 of vascular cell adhesion molecule (VCAM)-1 in buffer containing 15 μg ml−1 of the β7 activating antibody 2G3. (d) CTCL cells were cultured for 72 hours in the presence of 100 nM ATRA, 1 μM Bexarotene, or vehicle. Adhesion was the determined on immobilized MAdCAM-1-Fc in the absence or presence of the pan-integrin activator Mn2+.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

4. Figure 3
CCL25 activation of β7 integrin function is heightened in cutaneous T-cell lymphoma (CTCL) cells exposed to retinoids. (a) HuT78 cells exposed to 100 nM all-trans-retinoic acid (ATRA) (gray), 1 μM Bexarotene (black), or DMSO (white) for 72 hours were incubated with 5 μg ml−1 of soluble mucosal addressin cell adhesion molecule (MAdCAM)-1-Fc. Cell surface binding of MAdCAM-1-Fc was disclosed with anti-Fc antibodies via flow cytometry. (b) Soluble binding assays described in a were repeated with HuT78 cells pre-incubated with 1 μg ml−1 of CCL25.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

5. Figure 4
Augmented β7-mediated cutaneous T-cell lymphoma (CTCL) cell adhesion is dependent upon the presence of vitamin-A derivatives. (a) Adhesion of HuT78 cells exposed to the indicated doses of all-trans-retinoic acid (ATRA) and MJ cells exposed to various concentrations of Bexarotene for 72 hours was determined. (b) HuT78 and MJ cells were cultured with 100 nM ATRA (filled squares), 1 μM Bexarotene (filled triangles), or vehicle (open circles) for 72 hours. After the initial exposure, cells were washed with media to remove retinoids. Washed cells were then sub-cultured in media lacking retinoids for the indicated time (abscissa). (c) Vitamin or vitamin derivatives (100 nM) were incubated with HuT78 or MJ cells for 72 hours and adhesion was assessed. All assays utilized 0.75 μg ml−1 of immobilized addressin cell adhesion molecule (MAdCAM)-1-Fc.
Journal of Investigative Dermatology  , DOI: ( /jid )
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6. Figure 5
The vitamin D metabolite, 1,25(OH)2D3, inhibits retinoid action in cutaneous T-cell lymphoma (CTCL) cell adhesion. MJ cells were treated with vehicle (white), 100 nM Bexarotene (black), or in combination of increasing levels of 1,25-(OH)2D3 (gray), or 100 nM vitamin K2 (hatched) for 72 hours. Cells were then assessed for adhesion changes to addressin cell adhesion molecule (MAdCAM)-1-Fc. The impact 1,25(OH)2D3 was also determined with HuT78 cells and all-trans-retinoic acid (ATRA). All conditions included MnCl2 for integrin activation.
Journal of Investigative Dermatology  , DOI: ( /jid )
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7. Figure 6
Retinoids induce cutaneous T-cell lymphoma (CTCL) cell adhesion through retinoic acid receptor (RAR)/retinoid X receptor (RXR) nuclear receptor synergism. MJ or HuT78 cells were cultured for 48 hours in the presence of vehicle, single RAR or RXR agonists, or combination of the RAR and RXR agonists. The RAR agonist TTNPB was used at 5 nM with the RXR agonists, Bexarotene and SR11237, both being administered at 100 nM. Cell adhesion was determined on 0.75 μg ml−1 of MAdCAM-1-Fc. Absorption values were converted to adherent cells per well by using a standard curve. Significance reflects combined agents versus single agents.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions