Journal: Scientific Reports

Article Title: Distribution and postnatal development of chondroitin sulfate proteoglycans in the perineuronal nets of cholinergic motoneurons innervating extraocular muscles

doi: 10.1038/s41598-022-25692-3

Figure Lengend Snippet: Lectin and primary antibodies used.

Article Snippet: Reaction was visualized with donkey anti-goat IgG AlexaFluor 488 (Invitrogen; Eugene, OR, USA; RRID:AB_2534102) in PBS at RT for 1 h. ChAT labeling was coupled with one of the following histochemical or immunohistochemical reactions: N-acetylgalactosamin substituents of chondroitin sulfate proteoglycans were labeled with biotinylated WFA lectin (Sigma-Aldrich, St. Louis, MO, USA ; RRID:AB_2620171), dissolved in PBS containing 1% BSA for one overnight at 4 °C, visualized with Streptavidin AlexaFluor 555 (Invitrogen; RRID:AB_2571525) secondary reagent, incubated in PBS for 1 h at RT; or anti-aggrecan (rabbit; Merck-Millipore; RRID:AB_90460) diluted in PBS containing 3% NGS + 1% BSA at 4 °C overnight, then visualized with goat anti-rabbit AlexaFluor 555 (Invitrogen; RRID:AB_2535850) diluted in PBS at RT for 1 h; or anti-brevican (sheep; R&D Systems, Minneapolis, MN, USA; RRID:AB_2274793) in PBS containing 3% NDS + 1% BSA at 4 °C, overnight, then visualized with donkey anti-sheep AlexaFluor 555 (Invitrogen; RRID:AB_2535857) diluted in PBS at RT for 1 h; or anti-neurocan (sheep; R&D Systems; RRID:AB_2149717) diluted in PBS containing 3% NDS + 1% BSA at 4 °C, overnight, then visualized with donkey anti-sheep AlexaFluor 555 (Invitrogen; RRID:AB_2535857) diluted in PBS at RT for 1 h; or anti-versican (rabbit; Biorbyt, Cambridge, UK) diluted in PBS containing 3% NGS + 1% BSA at 4 °C for three overnights, then visualized with biotinylated goat anti-rabbit IgG (Vector Laboratories, Burlingame, CA, USA; RRID:AB_2313606) for 1 h at RT and StreptAvidin AlexaFluor 555 (Invitrogen; RRID:AB_2571525) at RT for 1 h, both diluted in PBS (Tables and ).

Techniques: Isolation, Derivative Assay, Recombinant, Sequencing

Journal: Scientific Reports

Article Title: Distribution and postnatal development of chondroitin sulfate proteoglycans in the perineuronal nets of cholinergic motoneurons innervating extraocular muscles

doi: 10.1038/s41598-022-25692-3

Figure Lengend Snippet: Fluorescent secondary antisera.

Article Snippet: Reaction was visualized with donkey anti-goat IgG AlexaFluor 488 (Invitrogen; Eugene, OR, USA; RRID:AB_2534102) in PBS at RT for 1 h. ChAT labeling was coupled with one of the following histochemical or immunohistochemical reactions: N-acetylgalactosamin substituents of chondroitin sulfate proteoglycans were labeled with biotinylated WFA lectin (Sigma-Aldrich, St. Louis, MO, USA ; RRID:AB_2620171), dissolved in PBS containing 1% BSA for one overnight at 4 °C, visualized with Streptavidin AlexaFluor 555 (Invitrogen; RRID:AB_2571525) secondary reagent, incubated in PBS for 1 h at RT; or anti-aggrecan (rabbit; Merck-Millipore; RRID:AB_90460) diluted in PBS containing 3% NGS + 1% BSA at 4 °C overnight, then visualized with goat anti-rabbit AlexaFluor 555 (Invitrogen; RRID:AB_2535850) diluted in PBS at RT for 1 h; or anti-brevican (sheep; R&D Systems, Minneapolis, MN, USA; RRID:AB_2274793) in PBS containing 3% NDS + 1% BSA at 4 °C, overnight, then visualized with donkey anti-sheep AlexaFluor 555 (Invitrogen; RRID:AB_2535857) diluted in PBS at RT for 1 h; or anti-neurocan (sheep; R&D Systems; RRID:AB_2149717) diluted in PBS containing 3% NDS + 1% BSA at 4 °C, overnight, then visualized with donkey anti-sheep AlexaFluor 555 (Invitrogen; RRID:AB_2535857) diluted in PBS at RT for 1 h; or anti-versican (rabbit; Biorbyt, Cambridge, UK) diluted in PBS containing 3% NGS + 1% BSA at 4 °C for three overnights, then visualized with biotinylated goat anti-rabbit IgG (Vector Laboratories, Burlingame, CA, USA; RRID:AB_2313606) for 1 h at RT and StreptAvidin AlexaFluor 555 (Invitrogen; RRID:AB_2571525) at RT for 1 h, both diluted in PBS (Tables and ).

Techniques: Plasmid Preparation