Table 3.

Description of 36 of 96 Significantly and Near-Significantly Modulated Genes. Fold Change vs Control for Each Gene on the PCR Panel. Modulation of Gene Expression by RFV3 vs Glycerin Negative Control Expressed as Fold Change (FC; Up- or Down-Regulation)

Gene Symbol	p-value*/Average Threshold Cycles	Fold Up- or Down-Regulation	RFV3 Near-Differentially and Differentially- Expressed Genes (Comments and References)
KRT1	0.011 ‡	−2.6	Keratin, type II cytoskeletal 1; Krt1 participates in an inflammatory network in murine keratinocytes.61
KRT5	0.024 ‡	1.8	Keratin, type II cytoskeletal 5; involved in proliferation and differentiation of stratified epithelial cells.62
KRT6B	0.014 ‡	2.1	Keratin, type II cytoskeletal 6B; KRT6/16/17 recognized as key early barrier alarmins and upregulation of these keratins alters proliferation, cell adhesion, migration and inflammatory features of KCs.63
KRT17	0.000 ‡	−2.6	Keratin, type I cytoskeletal 17; KRT6/16/17 recognized as key early barrier alarmins and upregulation of these keratins alters proliferation, cell adhesion, migration and inflammatory features of KCs.63
CASP1	0.050 ‡	2.0	Caspase-1; Thiol protease that cleaves IL-1 beta between an Asp and an Ala, releasing the mature cytokine which is involved in a variety of inflammatory processes. Important for defence against pathogens. Can also promote apoptosis. Converting enzyme involved in keratinocyte terminal differentiation and cornification.64
TIMP1	0.081 ‡	2.1	Metalloproteinase inhibitor 1; Metalloproteinase inhibitor that functions by forming one to one complexes with target metalloproteinases, such as collagenases, and irreversibly inactivates them by binding to their catalytic zinc cofactor.
FGF2	0.007 ‡	2.0	Fibroblast growth factor 2; Plays an important role in the regulation of cell survival, cell division, angiogenesis, cell differentiation and cell migration. Functions as potent mitogen in vitro. Can induce angiogenesis; Belongs to the heparin-binding growth factors family regulatory protein in aging skin remodeling and wound healing.
IL6	0.016 ‡	−2.5	Interleukin-6; Cytokine with a wide variety of biological functions. It is a potent inducer of the acute phase response. Has involvement in atopic dermatitis and psoriasis with interdependence to PPAR-α.65,67
SERPINE1	0.005 ‡	1.8	Plasminogen activator inhibitor 1; Serine protease inhibitor. This inhibitor acts as “bait” for tissue plasminogen activator, urokinase, protein C and matriptase-3/TMPRSS7.
FOXO1	0.005 ‡	1.9	Forkhead box protein O1; Transcription factor that is the main target of insulin signalling and regulates metabolic homeostasis in response to oxidative stress. Known as pro-apoptotic, keratinocyte-differentiating.
PLAUR	0.013 ‡	2.4	Urokinase plasminogen activator surface receptor; Acts as a receptor for urokinase plasminogen activator. Plays a role in localizing and promoting plasmin formation. Possible ECM remodeling.
HGF	0.022 †	−8.1	Hepatocyte growth factor; linked to melanocyte proliferation.68
HSF1	0.043 ‡	2.3	Heat shock factor protein 1; Function as a stress-inducible and DNA-binding transcription factor that plays a central role in the transcriptional activation of the heat shock response (HSR) – HSPs are known to be linked with PPAR activation
VEGFA	0.010 ‡	1.6	Vascular endothelial growth factor A; Growth factor active in angiogenesis, vasculogenesis and endothelial cell growth. Induces endothelial cell proliferation, promotes cell migration, inhibits apoptosis and induces permeabilization of blood vessels.
AQP3	0.030 ‡	2.4	Aquaporin-3; Water channel required to promote glycerol permeability and water transport across cell membranes. Acts as a glycerol transporter in skin and plays an important role in regulating SC (stratum corneum) and epidermal glycerol content. Involved in skin hydration, wound healing, and tumorigenesis. Known to be mediated by PPAR.67
CDH1	0.002 ‡	2.6	Cadherin-1; Cadherins are calcium-dependent cell adhesion proteins. Cadherins are involved in dermal-epidermal adhesion.
LCE1D	0.043 ‡	2.3	Late cornified envelope protein 1D; Precursors of the cornified envelope of the stratum corneum.
PANX1	0.052 ‡	2.4	Pannexin-1; Structural component of the gap junctions and the hemichannels. May play a role as a Ca(2+)-leak channel to regulate ER Ca(2+) homeostasis.
PDCD6	0.068 ‡	2.2	Programmed cell death protein 6.
CERS3	0.007 ‡	1.6	Ceramide synthase 3; Has (dihydro) ceramide synthesis activity with relatively broad substrate specificity, but a preference for C18:0 and other middle- to long-chain fatty acyl-CoAs (By similarity). It is crucial for the synthesis of very long-chain ceramides in the epidermis, to maintain epidermal lipid homeostasis and terminal differentiation.69 Ceramide synthesis is known to be mediated by PPAR.67
POT1	0.001 ‡	1.9	Protection of telomeres protein 1; Component of the telomerase ribonucleoprotein (RNP) complex that is essential for the replication of chromosome termini.
S100A8	0.051 ‡	3.3	Protein S100-A8; S100A8 is a calcium- and zinc-binding protein which plays a prominent role in the regulation of inflammatory processes and immune response. It can induce neutrophil chemotaxis and adhesion.
S100A9	0.044 ‡	−2.9	Protein S100-A9; S100A9 is a calcium- and zinc-binding protein which plays a prominent role in the regulation of inflammatory processes and immune response
SIRT1	0.082 ‡	1.8	NAD-dependent protein deacetylase sirtuin-1; NAD-dependent protein deacetylase that links transcriptional regulation directly to intracellular energetics and participates in the coordination of several separated cellular functions such as cell cycle, response to DNA damage, metabolism, apoptosis and autophagy. Loss of sirtuin 1 (SIRT1) disrupts skin barrier integrity and sensitizes mice to epicutaneous allergen challenge.70
SMAD2	0.053 ‡	1.7	Mothers against decapentaplegic homolog 2; Receptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. TGF-b signal transduction pathway regulator.
TERF2	0.094 ‡	1.6	Telomeric repeat-binding factor 2; Binds the telomeric double-stranded 5ʹ-TTAGGG-3ʹ repeat and plays a central role in telomere maintenance and protection against end-to-end fusion of chromosomes.
TFAM	0.049 ‡	2.1	Transcription factor A, mitochondrial; Binds to the mitochondrial light strand promoter and functions in mitochondrial transcription regulation. Mitochondrial transcription factor.
TFB2M	0.056 ‡	1.7	Dimethyladenosine transferase 2, mitochondrial; S-adenosyl-L-methionine-dependent methyltransferase which specifically dimethylates mitochondrial 12S rRNA at the conserved stem loop. Mitochondrial transcription factor.
TGFB1	0.005 ‡	1.8	Transforming growth factor beta-1; Multifunctional protein that controls proliferation, differentiation and other functions in many cell types. It positively and negatively regulates many other growth factors. Stimulates sustained production of collagen through the activation of CREB3L1. TGFB1 is a master regulator of skin anti-aging processes.
TLR2	0.003 ‡	2.9	Toll-like receptor 2; Co-operates with LY96 to mediate the innate immune response to bacterial lipoproteins and other microbial cell wall components.
TMEM135	0.006 ‡	2.1	Transmembrane protein 135; Involved in mitochondrial metabolism by regulating the balance between mitochondrial fusion and fission.
TMEM33	0.035 ‡	2.2	Transmembrane protein 33; Acts as a regulator of the tubular endoplasmic reticulum (ER) network.
TOLLIP	0.018 ‡	1.8	Toll-interacting protein; Component of the signalling pathway of IL-1 and Toll-like receptors. Inhibits cell activation by microbial products.
TXNIP	0.117 ‡	1.3	Thioredoxin-interacting protein; May act as an oxidative stress mediator by inhibiting thioredoxin activity or by limiting its bioavailability.
FLG	0.017 ‡	2.4	Filaggrin; Aggregates keratin intermediate filaments and promotes disulfide-bond formation among the intermediate filaments during terminal differentiation of mammalian epidermis. Filaggrin is known as an essential barrier protein mediated by PPAR.67
TNF	0.011 †	−13.7	Tumor necrosis factor; Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. Master regulator of inflammation. Linked with atopic dermatitis and psoriasis. PPAR-α exerting beneficial effects in atopic dermatitis.67

Notes: *p-value calculated by t-test calculated FRES vs glycerin negative control. ‡ expression of a particular gene was high (<30 cycles to visualize) in both control and the comparator. † gene’s average threshold cycle was high (>30) in either the control or the test sample, and was reasonably low in the other samples (<30).