CAF marker mRNA expression.

<p>Gene expression of CAF markers was studied using Q-PCR. (A) α-SMA, FSP1 and FAP expression ratios were significantly lower (P<0.01) in CAF-43 cells compared to FB-43 cells, but equal or higher in CAF-74 fibroblasts. (B) When grown as spheroids all fibroblasts downregulated α-SMA expression, but CAFs started to regain the expression at 72 h (P<0.05 in FB-43 vs. CAF-43 and in FB-74 vs. CAF-74) (C) FSP1 was downregulated in normal fibroblasts and in CAF-74 cells, but not in CAF-43 cells, (D) and FAP was upregulated in all cell lines going through nemosis, more so in CAFs (P<0.05 in 72 h CAF-43 when compared to 72 h FB-43 ). Columns: mean; error bars; SEM.</p

Senescence-associated beta-galactosidase activity of fibroblasts.

<p>Representative pictures of fibroblast monolayer cultures stained for SA-β-gal activity. FB-43 (A) and FB-74 (C) show little or no staining; CAFs are positive (B and D). Scale bar 200 µm. CAF-74 had significantly more (P<0.01) SA-β-gal cells compared to CAF-43 (E). Columns: mean; error bars; SEM.</p

Growth factor mRNA levels.

<p>Growth factor gene expression was studied using Q-PCR. (A and B) Both CAF cell lines had reduced expression of HGF/SF (P<0.05) and FGF7 (P<0.01) and all three growth factors were upregulated to a varying degree in nemosis (C – VEGF, D – HGF/SF and E – FGF7). Columns: mean; error bars; SEM.</p

Fibroblast growth in soft-agarose assay.

<p>Representative pictures of underlying fibroblast monolayer cultures. Spontaneous clustering (arrows) is seen in FB-43 and CAF-43 cells under the influence of paired SCC cells 43A (A and B) and 43B (C and D). In contrast, FB-74 (E and G) and CAF-74 (F and H) did not form spheroids. Scale bar 60 µm. The number of formed spheroids was calculated from the monolayer fibroblast cultures (I and J). CAF-43 cells formed significantly more spheroids than FB-43 cells (P<0.05). Columns: mean; error bars; SEM.</p

Soft-agarose assay scores.

<p>UT-SCC colony formation was studied with soft-agarose assay. All UT-SCC cells formed colonies in soft agarose, recurrent SCC (B and D) twice as many as primary SCC cells (A and C) (P<0.05). Normal fibroblasts increased the number of colonies of primary carcinomas cells and this was further augmented with CAF cells (P<0.05) (A and C). Recurrent SCC cell colony formation was inhibit with normal fibroblasts (P<0.05 in FB-74 compared to control) and restored to control level by CAFs (B and C). Columns: mean; error bars; SEM.</p

Primer sequences used in Q-PCR.

<p>Primer sequences used in Q-PCR.</p

Nemosis response in different fibroblast populations.

<p>Fibroblasts were grown as spheroids or monolayer for the time indicated. (A) FB-43 spheroids started to produce COX-2 after 48 hours and no α-SMA was produced, whereas CAF-43 cells (B) did not induce COX-2 but expressed α-SMA. Both FB-74 (C) and CAF-74 (D) produced α-SMA, but COX-2 was only induced in CAF-74 spheroids. All fibroblasts types expressed equal amounts of vimentin. (E) All UT-SCC cells expressed COX-2, but only 74A and 74B showed induced p53 levels.</p

Tula hantavirus isolate with the full-length ORF for nonstructural protein NSs survives for more consequent passages in interferon-competent cells than the isolate having truncated NSs ORF-2

<p><b>Copyright information:</b></p><p>Taken from "Tula hantavirus isolate with the full-length ORF for nonstructural protein NSs survives for more consequent passages in interferon-competent cells than the isolate having truncated NSs ORF"</p><p>http://www.virologyj.com/content/5/1/3</p><p>Virology Journal 2008;5():3-3.</p><p>Published online 11 Jan 2008</p><p>PMCID:PMC2253529.</p><p></p>specific primers. From up: results of RT-PCR assays with the primers specific for: TULV/Lodz S segment, TULV/Lodz M segment, TULV/Moravia S segment, and TULV/Moravia M segment

Tula hantavirus isolate with the full-length ORF for nonstructural protein NSs survives for more consequent passages in interferon-competent cells than the isolate having truncated NSs ORF-3

<p><b>Copyright information:</b></p><p>Taken from "Tula hantavirus isolate with the full-length ORF for nonstructural protein NSs survives for more consequent passages in interferon-competent cells than the isolate having truncated NSs ORF"</p><p>http://www.virologyj.com/content/5/1/3</p><p>Virology Journal 2008;5():3-3.</p><p>Published online 11 Jan 2008</p><p>PMCID:PMC2253529.</p><p></p> RNA was isolated. RT-PCR was done with the isolate-specific S- and M-primers. From top: results of RT-PCR assays with the primers specific for: TULV/Lodz S segment, TULV/Lodz M segment, TULV/Moravia S segment, and TULV/Moravia M segment

Antibody titration assays using AF-labeled or Eu-labeled protein L.

<p><b>A</b>) Anti-GST antibody titrated against Eu-labeled GST-VP1u and AF-labeled protein L. <b>B</b>) Anti-SA antibody titrated against Eu-labeled SA and AF-labeled protein L. <b>C</b>) Anti-SA antibody titrated against AF-labeled SA and Eu-labeled protein L. <b>D</b>) Anti-GST antibody titrated against AF-labeled GST-VP1u and Eu-labeled protein L. In all setups antibody concentrations were from 3.1 nM to 50 nM, and the antigen and protein concentration was constant (20 nM). Anti-GST antibody was used as a control for SA assays, and anti-SA for GST assays. The third line represents a background control with no antibody. The y-axis represents response counts obtained from Victor² fluorometer. The error bars represent ± standard deviation between parallel wells.</p