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The expression levels of 145- and 95-KDa TrkB (a). P75NTR (b), pro-BDNF and BDNF (c) as well as Akt1 and ERK2(d), Shc and N-Shc (e), NR1 and PLC-γ1(f), in 50 μg post-mitochondrial synaptosome-enriched fractions prepared from hippocampi and PFCX of P21 rats exposed to saline or cocaine in utero were compared by Western blotting. The blots were stripped and re-probed with anti-β-actin to validate equal loading. Densitometric quantification of blots revealed no discernible differences in 145- and 95-KDa TrkB, P75NTR, proBDNF and BDNF, Akt1, ERK2, Shc and N-Shc, NR1 and PLC-γ1 expression levels. n = 4. Data are mean ± s.e.m. of the ratio of 145-, 95-KDa TrkB, p75 NTR , proBDNF, BDNF, Akt1, ERK2, N-Shc, Shc, NR1 or PLC-γ1to β-actins optical intensities.

Journal: PLoS ONE

Article Title: Prenatal Cocaine Exposure Upregulates BDNF-TrkB Signaling

doi: 10.1371/journal.pone.0160585

Figure Lengend Snippet: The expression levels of 145- and 95-KDa TrkB (a). P75NTR (b), pro-BDNF and BDNF (c) as well as Akt1 and ERK2(d), Shc and N-Shc (e), NR1 and PLC-γ1(f), in 50 μg post-mitochondrial synaptosome-enriched fractions prepared from hippocampi and PFCX of P21 rats exposed to saline or cocaine in utero were compared by Western blotting. The blots were stripped and re-probed with anti-β-actin to validate equal loading. Densitometric quantification of blots revealed no discernible differences in 145- and 95-KDa TrkB, P75NTR, proBDNF and BDNF, Akt1, ERK2, Shc and N-Shc, NR1 and PLC-γ1 expression levels. n = 4. Data are mean ± s.e.m. of the ratio of 145-, 95-KDa TrkB, p75 NTR , proBDNF, BDNF, Akt1, ERK2, N-Shc, Shc, NR1 or PLC-γ1to β-actins optical intensities.

Article Snippet: Anti-phosphotyrosine (SC-508), TrkB (SC-119) -ERK2 (SC-154, SC-81457), -pT 202 / pY 204 -ERK (SC-136521), -pS 473 Akt (SC- 7985-R), -Akt1 (SC-65487), -Akt1/2/3 (SC-8312), -phospholipase C-γ1 (SC-7290), -Shc (SC-967), -N-Shc (SC-28833) -NR1 (SC-9058), -actin (SC-7210), -β-actin (SC-47778), -p75 NTR (SC-6189, SC-55467), -TRAF2 (SC-136999), -TRAF6 (SC-8409), tissue plasminogen activator (tPA) inhibitor were from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques: Expressing, Saline, In Utero, Western Blot

Prenatal cocaine exposure heightened TrkB-NMDAR interaction in hippocampi and PFCX as indicated by the Western blot detection of increased level of obligatory subunit of the NMDARs, NR1 in the anti-TrkB immunoprecipitates (top). Western blots were analyzed by densitometric quantification. The data are expressed as the ratios of NR1 optical intensity normalized by the optical intensity of total 145-KDa TrkB. n = 6 (3 males and 3 females). Data are means ± s.e.m. of the ratios of NR1 to 145-KDa TrkB optical intensities. *p < 0.01, compared to respective protein in the saline-treated group.

Journal: PLoS ONE

Article Title: Prenatal Cocaine Exposure Upregulates BDNF-TrkB Signaling

doi: 10.1371/journal.pone.0160585

Figure Lengend Snippet: Prenatal cocaine exposure heightened TrkB-NMDAR interaction in hippocampi and PFCX as indicated by the Western blot detection of increased level of obligatory subunit of the NMDARs, NR1 in the anti-TrkB immunoprecipitates (top). Western blots were analyzed by densitometric quantification. The data are expressed as the ratios of NR1 optical intensity normalized by the optical intensity of total 145-KDa TrkB. n = 6 (3 males and 3 females). Data are means ± s.e.m. of the ratios of NR1 to 145-KDa TrkB optical intensities. *p < 0.01, compared to respective protein in the saline-treated group.

Article Snippet: Anti-phosphotyrosine (SC-508), TrkB (SC-119) -ERK2 (SC-154, SC-81457), -pT 202 / pY 204 -ERK (SC-136521), -pS 473 Akt (SC- 7985-R), -Akt1 (SC-65487), -Akt1/2/3 (SC-8312), -phospholipase C-γ1 (SC-7290), -Shc (SC-967), -N-Shc (SC-28833) -NR1 (SC-9058), -actin (SC-7210), -β-actin (SC-47778), -p75 NTR (SC-6189, SC-55467), -TRAF2 (SC-136999), -TRAF6 (SC-8409), tissue plasminogen activator (tPA) inhibitor were from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques: Western Blot, Saline