qPCR validation of RNA-seq result.

<p>Quantitative PCR for (A) <i>gli1</i>, (B) <i>ptch1</i>, and (C) <i>hsd11b1</i> was performed on Hh ligand treated and Hedgehog ligand and SFE co-treated TRAMPC2 cells. Transcripts concentrations were normalized to control. * indicates p<0.05. In the lower figure, transcripts concentrations of (A) <i>gli1</i>, (B) <i>ptch1</i>, and (C) <i>hsd11b1</i> are represented by quantified sequencing reads, in the form of counts-per-million-reads.</p

Sutherlandia Extract alters genes in TRAMPC2 cells.

<p>(A) Differentially expressed genes in response to Sutherlandia extract treatment. Genes that are related to (B, C, D) are labeled. (B) Gene Ontology analysis of Sutherlandia responsive genes. (C, D) KEGG pathway analysis of Sutherlandia responsive genes.</p

Heat map of Sutherlandia Extract altered Hedgehog-signaling pathway target genes expression.

<p>TRAMPC2 cell were treated with either Hh-CM or co-treated with Hh-CM and 80μg/ml SFE. Over 50% of Hh-responsive genes were repressed by SFE treatment. Gene expression values were represented by Log2 transformed normalized RNA-seq reads (Log2 count-per-million-reads) and color coded.</p

<i>S</i>. <i>frutescens</i> (SF) Consumption Failed to Impact Circulating Cytokines and Chemokines in BALB/c Mice 24 hr Following an <i>in vivo</i> Challenge with <i>L</i>. <i>monocytogenes</i>.^<i>a</i>

<p><i>S</i>. <i>frutescens</i> (SF) Consumption Failed to Impact Circulating Cytokines and Chemokines in BALB/c Mice 24 hr Following an <i>in vivo</i> Challenge with <i>L</i>. <i>monocytogenes</i>.<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0160994#t002fn001" target="_blank">^<i>a</i></a></p

An Investigation into the Immunomodulatory Activities of <i>Sutherlandia frutescens</i> in Healthy Mice

<div><p><i>Sutherlandia frutescens</i> is a medicinal plant that has been traditionally used in southern Africa for cancers, infections, and inflammatory conditions. We recently published experiments demonstrating that an aqueous extract of <i>S</i>. <i>frutescens</i> possessed potent immune-stimulatory activity. This work was carried out with murine macrophages, an immune cell type that plays a pivotal role in host defense from infection and in shaping host inflammatory and immune responses. Here, we conducted a series of follow-up experiments to explore the impact of consuming <i>S</i>. <i>frutescens</i> on host response to bacterial challenge using healthy mice. We found that feeding mice a diet containing <i>S</i>. <i>frutescens</i> failed to significantly alter host response to systemic infection by either a gram-positive or gram-negative bacterium (i.e., <i>L</i>. <i>monocytogenes</i> and <i>E</i>. <i>coli</i>, respectively). In contrast to the <i>in vitro</i> observations, we found no evidence that <i>S</i>. <i>frutescens</i> consumption stimulated <i>in vivo</i> inflammatory responses; instead, consumption of <i>S</i>. <i>frutescens</i> tended to diminish <i>in vivo</i> inflammatory responses. Several possible reasons for this are discussed.</p></div

Changes in biochemical and hematological parameters over time in the combined Stage 1 and Stage 2 analysis <i>S</i>. <i>frutescens</i> 1,200 mg (N = 54) and placebo (N = 53).

<p>Abbreviations: HDL (high density lipoprotein); LDL (low density lipoprotein)</p><p>*P-value for interaction effect of groups over time.</p><p>Changes in biochemical and hematological parameters over time in the combined Stage 1 and Stage 2 analysis <i>S</i>. <i>frutescens</i> 1,200 mg (N = 54) and placebo (N = 53).</p

Baseline demographic and clinical characteristics.

<p>Baseline demographic and clinical characteristics.</p

Dietary <i>S</i>. <i>frutescens</i> Has No Impact on Spontaneous Activity (i.e., sickness behavior) of Mice After an Experimental Infection with <i>E</i>. <i>coli</i>.

<p>Conditions were similar to those described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0160994#pone.0160994.g003" target="_blank">Fig 3</a>, except in this experiment the mice were C57Bl/6 mice and the challenge consisted of an intraperitoneal injection with ~10⁸ cfu of <i>E</i>. <i>coli</i> (K-12 strain).</p

Impact of <i>S</i>. <i>frutescens</i> Consumption on Host Clearance of an Experimental Infection with <i>L</i>. <i>monocytogenes</i>.

<p>Female and male BALB/c (<i>L</i>. <i>monocytogenes</i>) mice were randomly assigned to one of three treatment groups: 0, 0.25, or 1.0% (wt/wt) <i>S</i>. <i>frutescens</i> in an AIN93G-type diet. After consuming diets for ~4 wks, all mice received an intravenous injection with ~10⁴ cfu of <i>L</i>. <i>monocytogenes</i> (EGD strain) in 0.2 mL of sterile, endotoxin-free phosphate-buffered saline. Liver and spleen samples were harvested three days after challenge and colony-forming units (<i>cfu</i>) were determined by a fluorescence-based microplate assay of tissue homogenates. Each symbol represents the data from a single mouse with the median and intra-quartile range shown.</p

Impact of <i>S</i>. <i>frutescens</i> Consumption on Host Clearance of an Experimental Infection with <i>E</i>. <i>coli</i>.

<p>Weanling female and male C57Bl/6 mice were randomly assigned to one of three treatment groups: 0, 0.25, or 1.0% (wt/wt) <i>S</i>. <i>frutescens</i> in an AIN93G-type diet. After consuming diets for ~4 wks, all mice were injected intraperitoneally with ~10⁸ colony-forming units (<i>cfu</i>) of <i>E</i>. <i>coli</i> (K-12 strain) in 1 mL of sterile PBS. Liver and spleen samples were harvested three days after challenge and colony-forming units (<i>cfu</i>) were determined by a fluorescence-based microplate assay of tissue homogenates. Each symbol represents the data from a single mouse with the median and intra-quartile range shown.</p