Journal: Cell Reports Methods

Article Title: BTB BCL6 dimers as building blocks for reversible drug-induced protein oligomerization

doi: 10.1016/j.crmeth.2022.100193

Figure Lengend Snippet:

Article Snippet: anti-p44/42 MAPK (Erk1/2) , Cell Signaling Technology , #9102.

Techniques: Virus, Recombinant, Software, Transfection, Cell Culture, Expressing, Electron Microscopy

Journal: Scientific Reports

Article Title: BMP7 retards peripheral myelination by activating p38 MAPK in Schwann cells

doi: 10.1038/srep31049

Figure Lengend Snippet: ( A ) The SCs were transfected with SMAD4 siRNAs for 72 h. The mRNA levels of SMAD4 were analyzed by q-PCR. The control cells were transfected with negative control (NC) siRNA. ( B ) The SCs were transfected with SMAD4 siRNA-2 or NC siRNA for 48 h, and then the cells were treated with BMP7 (50 ng/ml) for additional 24 h. The total cell lysates were prepared for measuring SMAD and p38 MAPK by Western blot. ( C ) Densitometric quantification of the immunoblot data in ( B ). ( D,E ) The SCs were transfected with SMAD4 siRNA-2 or NC siRNA. 48 h post-transfection, the cells were treated with BMP7 (50 ng/ml) and cAMP (1 mM) as indicated for additional 24 h. The mRNA levels of Krox20, Oct6 and Pmp22 were analyzed by q-PCR ( D ). The protein levels of Oct6, Krox20 and PMP22 were analyzed by Western blot ( E ). ( F ) Densitometric quantification of the immunoblot data in ( E ). Actin was used as the loading control. Data are representative of three independent experiments. Three samples were employed for q-PCR and Western blot analysis. Error bars are ± SEM. n.s. means no significance. *p < 0.05, **p < 0.01, Student’s t test for ( A ); one-way ANOVA with Bonferroni’s post hoc testing for ( C , D , F ).

Article Snippet: Rabbit anti-phospho-Erk, rabbit anti-Erk, rabbit anti-phospho-JNK, rabbit anti-JNK, rabbit anti-phospho-p38 MAPK, rabbit anti-p38 MAPK, rabbit anti-phospho-SMAD1/5/8, rabbit anti-SMAD5, rabbit anti-phospho-ATF2, rabbit anti-ATF2 and rabbit anti-Tubulin antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Transfection, Negative Control, Western Blot

Journal: Scientific Reports

Article Title: BMP7 retards peripheral myelination by activating p38 MAPK in Schwann cells

doi: 10.1038/srep31049

Figure Lengend Snippet: ( A ) The SCs were treated with BMP7 (50 ng/ml) and cAMP (1 mM) as indicated and MAPKs were analyzed by Western blot. ( B ) Densitometric quantification of the immunoblot data in ( A ). ( C ) The SCs were treated with BMP7 (50 ng/ml) and sb203580 (10 μM) as indicated for 24 h. The protein levels of p-p38 MAPK and p-ATF2 were analyzed by Western blot. ( D ) Densitometric quantification of the immunoblot data in ( C ). ( E ) p38 MAPK inhibition by sb203580 restores the decreases in myelin gene expression induced by BMP7. The primary rat SCs treated with BMP7 (50 ng/ml), cAMP (1 mM) and sb203580 (10 μM) as indicated and the mRNA levels of Oct6, Krox20, MBP, MPZ and PMP22 was measured by q-PCR. ( F ) p38 MAPK inhibition by sb203580 diminishes the attenuation effects of BMP7 on Krox20, Oct6 and PMP22 protein levels. The SCs were treated as indicated and the protein levels of Krox20 and Oct6 were analyzed by Western blot. ( G ) Densitometric quantification of the immunoblot data in ( F ). Actin was used as the loading control. Data are representative of three independent experiments. Three samples were employed for q-PCR and Western blot analysis. Error bars are ± SEM. n.s. means no significance. *p < 0.05, **p < 0.01, one-way ANOVA with Bonferroni’s post hoc testing.

Article Snippet: Rabbit anti-phospho-Erk, rabbit anti-Erk, rabbit anti-phospho-JNK, rabbit anti-JNK, rabbit anti-phospho-p38 MAPK, rabbit anti-p38 MAPK, rabbit anti-phospho-SMAD1/5/8, rabbit anti-SMAD5, rabbit anti-phospho-ATF2, rabbit anti-ATF2 and rabbit anti-Tubulin antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Western Blot, Inhibition, Expressing

Journal: Scientific Reports

Article Title: BMP7 retards peripheral myelination by activating p38 MAPK in Schwann cells

doi: 10.1038/srep31049

Figure Lengend Snippet: ( A , B ) The primary rat SCs were transfected with Mapk14 siRNAs for 72 h. The mRNA levels of Mapk14 ( A ) or the protein levels of p38 MAPK ( B ) were analyzed by q-PCR and Western blot, respectively. The control cells were transfected with negative control (NC) siRNA. ( C , D ) The primary rat SCs were transfected with Mapk14 siRNA-3 or NC siRNA. 48 h post-transfection, the cells were treated with cAMP (1 mM) and BMP7 (50 ng/ml) as indicated. The mRNA levels ( C ) or protein levels ( D ) of Oct6, Krox20 and Pmp22 were measured. Tubulin was used as the loading control. ( E ) Densitometric quantification of the immunoblot data in ( D ). Data are representative of three independent experiments. Three samples were employed for q-PCR and Western blot analysis. Error bars are ± SEM. n.s. means no significance. *p < 0.05, **p < 0.01, Student’s t test for ( A , B ); one-way ANOVA with Bonferroni’s post hoc testing for ( C , E ).

Article Snippet: Rabbit anti-phospho-Erk, rabbit anti-Erk, rabbit anti-phospho-JNK, rabbit anti-JNK, rabbit anti-phospho-p38 MAPK, rabbit anti-p38 MAPK, rabbit anti-phospho-SMAD1/5/8, rabbit anti-SMAD5, rabbit anti-phospho-ATF2, rabbit anti-ATF2 and rabbit anti-Tubulin antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Transfection, Western Blot, Negative Control

Journal: Scientific Reports

Article Title: BMP7 retards peripheral myelination by activating p38 MAPK in Schwann cells

doi: 10.1038/srep31049

Figure Lengend Snippet: ( A ) The cultured primary rat SCs were treated as indicated and the c-Jun mRNA levels were detected by q-PCR. ( B ) The protein levels of p38 MAPK, c-Jun and PMP22 in developing rat sciatic nerves isolated from E18, P2, P4, P6, P8, P10 and adult (Ad) rats were analyzed by Western blot. n = 4 for each group. Actin was used as the loading control. Data are representative of three independent experiments. Three samples were employed for q-PCR. Error bars are ± SEM. **p < 0.01, one-way ANOVA with Bonferroni’s post hoc testing.

Article Snippet: Rabbit anti-phospho-Erk, rabbit anti-Erk, rabbit anti-phospho-JNK, rabbit anti-JNK, rabbit anti-phospho-p38 MAPK, rabbit anti-p38 MAPK, rabbit anti-phospho-SMAD1/5/8, rabbit anti-SMAD5, rabbit anti-phospho-ATF2, rabbit anti-ATF2 and rabbit anti-Tubulin antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Cell Culture, Isolation, Western Blot

Journal: Scientific Reports

Article Title: BMP7 retards peripheral myelination by activating p38 MAPK in Schwann cells

doi: 10.1038/srep31049

Figure Lengend Snippet: ( A , B ) Recombinant BMP7 concentrations in blood ( A ) and sciatic nerves ( B ) after BMP7 injection. ( C ) The mRNA levels of Oct6, Krox20, Pmp22, Mbp and Mpz in sciatic nerves were analyzed by q-PCR. ( D ) The protein levels of p-p38 MAPK, p38 MAPK, Krox20, MPZ and PMP22 were analyzed by Western blot. ( E , F ) Analysis of sciatic nerves from vehicle- or BMP7-treated newborn rats by electron microscopy. Scale bar represents 5 μm ( E ) and 0.2 μm ( F ). n = 5 for each group. Error bars are ± SEM. **p < 0.01, Student’s t test.

Article Snippet: Rabbit anti-phospho-Erk, rabbit anti-Erk, rabbit anti-phospho-JNK, rabbit anti-JNK, rabbit anti-phospho-p38 MAPK, rabbit anti-p38 MAPK, rabbit anti-phospho-SMAD1/5/8, rabbit anti-SMAD5, rabbit anti-phospho-ATF2, rabbit anti-ATF2 and rabbit anti-Tubulin antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Recombinant, Injection, Western Blot, Electron Microscopy

Journal: The Journal of comparative neurology

Article Title: Cortical projections to the two retinotopic maps of primate pulvinar are distinct

doi: 10.1002/cne.24515

Figure Lengend Snippet: Distributions of labeled neurons in visual cortex after injections of CTB into the retinotopic maps in the lateral and inferior pulvinar in two galagos. a-b) Receptive fields for multiunit recordings in the dorsal and ventral maps in the pulvinar of galagos 16–25 and 16–26. The receptive fields are drawn on a depiction of the contralateral lower visual quadrant with 0° corresponding to area centralis of the retina. Receptive fields drawn in blue correspond to retinotopic mapping done using a tungsten electrode. Those fields drawn in red and green were obtained at the dorsal and ventral map injection sites, respectively. Indicated values are the cortical depths at which the circumscribed receptive fields were recorded. c-d) Coronal sections stained for CO overlaid on adjacent sections stained fluorescently for the injected CTB. Injection sites for the dorsal (red) and ventral (green) maps are shown. Scale bar is 1mm, e-f) The locations of populations of labeled neurons projecting to the dorsal map (red) and ventral map (green) in the coronal plane projected at a 45° angle onto the reconstructed dorsolateral surface of cortex. Borders of areas 17, 18, and MT indicated in black. Blue lines indicate tissue blocking cuts. Dashed grey lines indicate every ninth section of tissue. Scale bar is 1cm.

Article Snippet: Parts of the dorsal and ventral retinotopic maps were carefully mapped with single tungsten electrodes (FHC, Inc., ME) at stereotaxic coordinates A-P: 3 mm, M-L: 5.5 mm, using visually evoked potentials in response to a simple stimulus consisting of a spot of light containing a crosshair pattern.

Techniques: Labeling, Injection, Staining, Blocking Assay