Journal: Journal of Biological Chemistry

Article Title: Identification of Radil as a Ras binding partner and putative activator

doi: 10.1016/j.jbc.2021.100314

Figure Lengend Snippet: Figure 3. Radil interacts with active Ras proteins. A, HEK293T cells were transfected with the plasmid expressing Flag-tagged HRas (WT), constitutively active HRas (V12), or dominant-negative HRas (N17) for 24 h, after which cell lysates were immunoprecipitated with the anti-Flag antibody. Flag immu- noprecipitates, along with lysate inputs, were blotted for Radil and FLAG. The amount of Radil interacting with various forms of KRas was quantified via densitometry. Normalized values equal the density of Ras-bound Radil/density of total Radil/fold density of IP Flag. B, computer modeling structures of KRas/Radil interaction with or without GTP binding. C, HEK293T cells were transfected with plasmids expressing either WT or constitutively active forms of Ras (V12) proteins as indicated for 24 h, after which cell lysates were immunoprecipitated with the anti-Flag antibody. Flag immunoprecipitates, along with lysate inputs, were blotted for Radil and Flag. The amount of Radil interacting with various forms of KRas was quantified via densitometry. Normalized values equal the density of Ras-bound Radil/density of total Radil/Fold density of IP Flag. D, HEK293T cells were transfected with plasmids expressing GFP- KRas and/or Flag-Radil for 24 h, after which cell lysates were immunoprecipitated with the anti-Flag antibody. Flag immunoprecipitates, along with lysate inputs, were blotted for Flag, GFP, Rap1, and ArhGAP29.

Article Snippet: Polyclonal Radil and ArhGap29 antibody were purchased from ProteinTech.

Techniques: Transfection, Plasmid Preparation, Expressing, Dominant Negative Mutation, Immunoprecipitation, Binding Assay

Journal: PLoS ONE

Article Title: Allergen-Induced Dermatitis Causes Alterations in Cutaneous Retinoid-Mediated Signaling in Mice

doi: 10.1371/journal.pone.0071244

Figure Lengend Snippet: Systemic and topical OVA sensitization results in inflammation, disturbs epidermal barrier homeostasis, and induces PPARδ target gene expression in skin.

Article Snippet: FABP5 protein levels were determined in protein lysates prepared from whole mouse skin according to the protocol indicated in using the rabbit FABP5 polyclonal antibody purchased from ProteinTech (Chicago, IL).

Techniques: Targeted Gene Expression, Binding Assay

Journal: PLoS ONE

Article Title: Allergen-Induced Dermatitis Causes Alterations in Cutaneous Retinoid-Mediated Signaling in Mice

doi: 10.1371/journal.pone.0071244

Figure Lengend Snippet: ( a ) IL-4 serum levels after systemic with or without additional topical OVA sensitization (n = 8). ( b ) ATRA levels in mouse skin determined by HPLC MS-MS method upon systemic (i.p.) and systemic plus topical (i.p.+e.c.) OVA sensitization (n = 3/group). ( c ) Ratio of Fabp5 vs. Crabp2 expression in the skin of OVA-treated mice (n = 6/group) compared to control mice (PBS i.p.). Data are presented as mean values ± SEM. Statistical significance ( p ) is based on one-way ANOVA followed by Tukey’s multiple comparison test for gene expression results and ELISA data. For HPLC MS-MS results, significance was determined using Student’s t -test.

Article Snippet: FABP5 protein levels were determined in protein lysates prepared from whole mouse skin according to the protocol indicated in using the rabbit FABP5 polyclonal antibody purchased from ProteinTech (Chicago, IL).

Techniques: Tandem Mass Spectroscopy, Expressing, Control, Comparison, Gene Expression, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Allergen-Induced Dermatitis Causes Alterations in Cutaneous Retinoid-Mediated Signaling in Mice

doi: 10.1371/journal.pone.0071244

Figure Lengend Snippet: ( a ) Fabp5 protein levels in the skin of mice with allergen-induced dermatitis. 150 µg proteins were loaded per lane and beta-actin was used as control for even protein loading. ( b ) Immunohistochemical analysis of Fabp5 protein expression in five-micrometer back skin sections of OVA-sensitized mice. ( c ) ATRA-induced nuclear receptor-mediated signaling pathways depending on the predominant cellular transport protein.

Article Snippet: FABP5 protein levels were determined in protein lysates prepared from whole mouse skin according to the protocol indicated in using the rabbit FABP5 polyclonal antibody purchased from ProteinTech (Chicago, IL).

Techniques: Control, Immunohistochemical staining, Expressing, Protein-Protein interactions