N. lugens 5th instar nymphs were immunized with a microinjection of heat-killed Escherichia coli K12 or Bacillus subtilis (500 cells per nymph, respectively) using the FemtoJet Microinjection System (Eppendorf-Nethler-Hinz, Hamburg, Germany). Nymphs were collected at different time points (6, 12 and 24 hours) after infection for the bacteria-induced expression experiment as previously described
[13 (link)].
For studies of tissue-specific expression, the N. lugens were dissected under a Leica S8AP0 stereomicroscope (Leica Microsystems GmbH, Wetzlar, Germany). The fat body, midgut, salivary gland, male reproductive system, ovary and carcass (the remaining body) were isolated and quickly washed in cold diethylpyrocarbonate (DEPC)-treated NaCl/Pi solution (137 mM NaCl, 2.68 mM KCl, 8.1 mM Na2HPO4, 1.47 mM KH2PO4, pH 7.4) and immediately frozen at -80°C as previously described
[2 (link), 13 (link)]. For each tissue, more than 300 N. lugens individuals were dissected and used for RNA extractions.
[13 (link)].
For studies of tissue-specific expression, the N. lugens were dissected under a Leica S8AP0 stereomicroscope (Leica Microsystems GmbH, Wetzlar, Germany). The fat body, midgut, salivary gland, male reproductive system, ovary and carcass (the remaining body) were isolated and quickly washed in cold diethylpyrocarbonate (DEPC)-treated NaCl/Pi solution (137 mM NaCl, 2.68 mM KCl, 8.1 mM Na2HPO4, 1.47 mM KH2PO4, pH 7.4) and immediately frozen at -80°C as previously described
[2 (link), 13 (link)]. For each tissue, more than 300 N. lugens individuals were dissected and used for RNA extractions.
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