Cell growth of recombinant <i>E. coli</i> JM109 strains in the presence of 4% (v/v) cyclohexane.

<p>The recombinant strains were cultured at 37°C and cyclohexane (CH) was added when OD₆₆₀ reached 0.2 (indicated by the arrow). □, pQE-80L (as the control); ▾, pQE-<i>mmsB</i>; •, pQE- <i>tsf</i>; △, pQE-<i>PSEEN0851</i>.</p

Colony formation of <i>E. coli</i> JM109 strains over-expressing <i>mmsB</i>, <i>tsf</i>, and <i>PSEEN0851</i> on LBGMg agar overlaid with decalin after 24 h incubation at 37°C. JM109 carrying empty pQE-80L was used as the control.

<p>Colony formation of <i>E. coli</i> JM109 strains over-expressing <i>mmsB</i>, <i>tsf</i>, and <i>PSEEN0851</i> on LBGMg agar overlaid with decalin after 24 h incubation at 37°C. JM109 carrying empty pQE-80L was used as the control.</p

Five high-abundance protein spots identified by MALDI-TOF/TOF.

a<p>The expression level of <i>P. putida</i> JUCT1 grown in nutrient medium without solvent.</p>b<p>The expression level of <i>P. putida</i> JUCT1 grown in the presence of 60% (v/v) cyclohexane.</p

Effect of organic solvents on cell growth of <i>P. putida</i> JUCT1 (open bar) and its parent strain JUCS (gray bar).

<p>The strains were initially grown in nutrient medium at 37°C till OD₆₆₀ reached 0.2, and then 60% (v/v) organic solvent was added for further incubation of 5 h.</p

The oxidation of cyclohexane by <i>E. coli</i> strains.

<p>Incubation condition: 37°C for 8 h with cyclohexane (1%, w/v).</p

SDS-PAGE analysis of expression of <i>mmsB</i>, <i>PSEEN0851</i>, and <i>tsf</i> in <i>E. coli</i> JM109.

<p>The transformants were grown in LB at 37°C and induced by 1 mM IPTG. Lanes: M, molecular mass marker; 1, pQE-80L (as control); 2, pQE-<i>mmsB</i>; 3, pQE-<i>PSEEN0851</i>; 4, pQE-<i>tsf</i>. Arrowheads indicate the locations of recombinant proteins.</p

MALDI-TOF/TOF analysis of peptides from 5 high-abundance proteins.

a<p>Number of peptides identified for individual protein in MALDI-TOF/TOF analysis.</p>b<p>Number of unique peptides identified for individual protein.</p>c<p>Total amino acid sequence of peptides identified for individual protein.</p>d<p>Percentage of amino acid sequence covered by peptides of individual protein.</p

BmTCTP-mediated phagocytosis by silkworm hemoctyes.

<p>Internalized red fluorescence-labeled latex beads (A) and FITC-labeled bacteria (B, C) pre-coated by BmTCTP or BSA in hemocytes which were stained with DAPI for nucleus in blue or DiO for cytoplasmic membrane in green (A) were subjected to analysis by fluorescence microscopy. Experiment was repeated three times and representative image were shown here. The percentage of engulfed targets was obtained by multiplying the % of phagocytosing cells with the mean number of internalized beads/bacteria. Each histogram corresponds to the mean value of samples from 5–8 larvae (± S.D.). Significant differences between BmTCTP and BSA treatment are marked by asterisks (*<i>p</i><0.05, **<i>p</i><0.01).</p

Expression and localization of BmTCTP in the silkworm midgut.

<p>A. The existence of BmTCTP in midgut lumen, peritrophic membrane, gut juice and hemolymph of the silkworm was analyzed by Western blotting. Equal amount of protein (20 µg) extracted from each tissue was subjected to SDS-PAGE analysis followed with immunoblotting analysis. B. The expression of BmTCTP in midgut epithelial cells was analyzed by immunohistochemistry. The nucleus was stained with DAPI, and BmTCTP was stained with anti-BmTCTP polyclonal antibody followed with FITC-conjugated secondary antibody, rabbit pre-immune serum was used as negative control. Gu, gut lumen; Ep, epithelia. C. The expression level of BmTCTP in midgut lumen after oral infection of <i>B. bombyseptieus</i> or <i>S. marcescens</i> at indicated time point was examined by Western blotting, then normalized to the amount of BmTCTP level at 0 h and annotated as mean fold increase (± S.D.). Experiment was repeated three times and representative images were shown here.</p

Induction of anti-microbial peptides mRNA expression and activation of ERK signaling by BmTCTP.

<p>qRT-PCR analysis of the mRNA levels of <i>CecropinA1</i> and <i>Attacin</i> (A), <i>Tube</i>, <i>Pelle</i>, <i>FADD</i> and <i>Dredd</i> (B) in BmNs cells under BmTCTP or BSA treatment. In the presence of MEK inhibitor U0126 or PD098059, the mRNA levels of <i>CecropinA1</i> and <i>Attacin</i> after BmTCTP treatment were examined at different time points (C) or at different concentrations of inhibitors (D) by qRT-PCR. E. qRT-PCR analysis of the mRNA levels of <i>ERK</i>, <i>CecropinA1</i> and <i>Attacin</i> under BmTCTP treatment at indicated time points in dsRNA-transfected cells. F. ERK phosphorylation was examined at indicated time points after BmTCTP treatment, the phosphorylation intensity was normalized to the protein level of Tubulin and annotated as mean fold increase (± S.D.) over the normalized phosphorylation intensity at the 0 min. G. Membrane protein extracted from BmNs cells under BmTCTP treatment at indicated time points was immunoblotted with anti-phosphotyrosine antibody 4G10. The mRNA levels of all target molecules measured by qRT-PCR were normalized to the internal control and represented as mean ± S.D.</p