Journal:

Article Title: Control of megakaryocyte-specific gene expression by GATA-1 and FOG-1: role of Ets transcription factors

doi: 10.1093/emboj/cdf527

Figure Lengend Snippet: Fig. 5. Selective binding of Fli-1 to the αIIb Ets element. (A) αIIb and M1α Ets elements bind distinct Ets proteins. EMSA using nuclear extracts from NIH-3T3 cells. Note that there is little or no cross-competition between the αIIb and M1α Ets elements. (B) Nuclear extracts from Y10 cells (left panel) and fetal liver-derived primary megakaryocytes (right panel) were used. Anti-Fli-1 (αFli-1), but not control (ctr), antibodies reacted with a band (thick arrow), resulting in a supershift (thin arrow).

Article Snippet: A 2 µg aliquot of the indicated antibodies: anti-FLI-1 (C19), anti-Ets-1 (N-276), anti-Ets-2 (C-20) and anti-PU.1 (T-21) (Santa Cruz Biotechnology) was added to nuclear extracts for 20 min prior to the addition of the probe.

Techniques: Binding Assay, Derivative Assay

Journal:

Article Title: Control of megakaryocyte-specific gene expression by GATA-1 and FOG-1: role of Ets transcription factors

doi: 10.1093/emboj/cdf527

Figure Lengend Snippet: Fig. 6. Occupancy of the αIIb promoter by Fli-1 in vivo. ChIP assays using Y10 cells, rabbit and mouse anti-Fli-1 antibodies or isotype-matched control antibodies. Primer pairs were the same as in Figure 2. The GAPDH gene served as control.

Article Snippet: A 2 µg aliquot of the indicated antibodies: anti-FLI-1 (C19), anti-Ets-1 (N-276), anti-Ets-2 (C-20) and anti-PU.1 (T-21) (Santa Cruz Biotechnology) was added to nuclear extracts for 20 min prior to the addition of the probe.

Techniques: In Vivo

Journal:

Article Title: Control of megakaryocyte-specific gene expression by GATA-1 and FOG-1: role of Ets transcription factors

doi: 10.1093/emboj/cdf527

Figure Lengend Snippet: Fig. 7. Fli-1 mediates GATA-1/FOG-1 synergy on the αIIb promoter. Inset: schematic of the assay. The reporter construct contained 100 bp of αIIb promoter upstream region in which a GAL4-binding site was substituted for the Ets element. GAL4 fusion constructs were co- expressed as indicated.

Article Snippet: A 2 µg aliquot of the indicated antibodies: anti-FLI-1 (C19), anti-Ets-1 (N-276), anti-Ets-2 (C-20) and anti-PU.1 (T-21) (Santa Cruz Biotechnology) was added to nuclear extracts for 20 min prior to the addition of the probe.

Techniques: Construct, Binding Assay

Journal: Journal of Inflammation (London, England)

Article Title: Effects of long non-coding RNA Gm14461 on pain transmission in trigeminal neuralgia

doi: 10.1186/s12950-019-0231-1

Figure Lengend Snippet: Effect of Gm14461 expression on MWT and expression of Gm14461, CGRP, P2X3 receptor, P2X7 receptor, TNF-α, IL-1β, and IL-6 in TN mice. C57BL/6 J mice were randomly divided into six groups ( n = 8/group): Sham, TN, TN + Scramble siRNA, TN + si-Gm14461, TN + Vector, and TN + Gm14461 group. a Gm14461 expression in mouse TGs was detected by qRT-PCR. b The MWT was measured to assess the nociception of mice at different times after CCI-ION surgery (0, 1, 3, 5, 7, 9, 11, 13, 15 d). * P < 0.05: Sham vs. TN; # P < 0.05 Scramble siRNA vs. si-Gm14461; $ P < 0.05 Vector vs. Gm14461. c The protein levels of CGRP, P2X3 receptor, and P2X7 receptor were examined using western blot. d-f The mRNA levels of TNF-α ( d ), IL-1β (e), and IL-6 (f) in mouse TGs were detected by qRT-PCR. Data are presented as mean ± SD. * P < 0.05 vs. Sham; # P < 0.05 vs. Scramble siRNA; $ P < 0.05 vs. Vector

Article Snippet: The membrane was then blocked with 5% non-fat dry milk and incubated with the primary antibodies against the CGRP, P2X3, and P2X7 (1:1000; all purchased from Santa Cruz Biotechnology).

Techniques: Expressing, Plasmid Preparation, Quantitative RT-PCR, Western Blot

Journal: Journal of Inflammation (London, England)

Article Title: Effects of long non-coding RNA Gm14461 on pain transmission in trigeminal neuralgia

doi: 10.1186/s12950-019-0231-1

Figure Lengend Snippet: Effect of Gm14461 expression on the expression of CGRP, P2X3 receptor, and P2X7 receptor in primary TGNs. a The primary mouse TGNs were isolated from C57BL/6 J mice and treated with TNF-α (10 ng/mL), IL-1β (25 ng/mL) and IL-6 (25 ng/mL) for 24 h. Gm14461 expression in TGNs was examined by qRT-PCR. * P < 0.05 vs. Control. b Western blot analysis of protein levels of CGRP, P2X3 receptor, and P2X7 receptor in primary TGNs transfected with scramble siRNA, si-Gm14461, empty vector, and pcDNA3.1-Gm14461 in the stimulation of TNF-α (10 ng/mL) for 24 h. c The qualifications of western blots in ( b ). Data are presented as mean ± SD. N = 3. * P < 0.05 vs. Control; # P < 0.05 vs. TNF-α + Scramble siRNA; $ P < 0.05 vs. TNF-α + Vector

Article Snippet: The membrane was then blocked with 5% non-fat dry milk and incubated with the primary antibodies against the CGRP, P2X3, and P2X7 (1:1000; all purchased from Santa Cruz Biotechnology).

Techniques: Expressing, Isolation, Quantitative RT-PCR, Control, Western Blot, Transfection, Plasmid Preparation

Journal: The Journal of Cell Biology

Article Title: Merlin controls the repair capacity of Schwann cells after injury by regulating Hippo/YAP activity

doi: 10.1083/jcb.201606052

Figure Lengend Snippet: Merlin-null nerves show a transient hypomyelination and slight increase in SC numbers. (A and B) Transmission EM pictures of P6 sciatic nerves from control (NF2 +/+ ) and Merlin-null (NF2 −/− ) animals. (C and D) Transmission EM pictures of P60 sciatic nerves from control (C) and Merlin-null (D) animals. (A–D) n = 3 mice. Bars, 10 µm. (E). Western blot of control and Merlin-null nerves at P6. n = 3 pools; 1 pool = 3 nerves from three mice for each genotype. β-Tubulin was used as a loading control. (F) Counts of Hoechst-stained nuclei from transverse cryostat sections of P6 and P72 sciatic nerves showing increases in cell number in Merlin-null nerves. For NF2 +/+ , n = 3; for NF2 −/− , n = 4. (G) Small but nonsignificant increase in G ratio in P60 Merlin-null nerves as compared with control. n = 3 mice. (F and G) Two-sided two-sample Student’s t test: **, P ≤ 0.004 (P6; F); **, P ≤ 0.005 (P72; F); P = 0.1 (G). Data are presented as means ± SEM.

Article Snippet: The following primary antibodies were used for Western blotting: myelin protein zero (1:1,000; SAB2500665; Sigma-Aldrich), MBP (1:2,000; sc-13912; Santa Cruz Biotechnology, Inc.), Krox20 (1:500; PRB-236P-100; Covance), β-tubulin (1:2,000; sc-134229; Santa Cruz Biotechnology, Inc.), cJun (1:1,000; 9165; Cell Signaling Technology), p-cJun (1:500; 9261; Cell Signaling Technology), p-Erk1/2 (1:2,000; 9101; Cell Signaling Technology), p-p38 (1:500; 4631; Cell Signaling Technology), YAP/TAZ (1:1,000; 8418; Cell Signaling Technology), TAZ (1:1,000; 4883; Cell Signaling Technology), YAP (1:1,000; 14074; Cell Signaling Technology), connective tissue growth factor (1:1,000; ab6992; Abcam), collagen IV (1:1,000; ab6586; Abcam), MCP1 (1:500; 500-P113; PeproTech), active β-catenin (1:1,000; 05–665; EMD Millipore), N-cadherin (1:1,000; ab76057; Abcam), E-cadherin (1:1,000; PharMingen Clone 36 610181; BD), p-Akt (1:2,000; 4060; Cell Signaling Technology), and vinculin (1:1,000; 4650; Cell Signaling Technology).

Techniques: Transmission Assay, Control, Western Blot, Staining