Journal:

Article Title: Cellular Sources of Transforming Growth Factor-? Isoforms in Early and Chronic Radiation Enteropathy

doi:

Figure Lengend Snippet: Intestinal wall immunoreactivity for TGF-β1 (anti-CC), TGF-β2, and TGF-β3 2 weeks (left) and 26 weeks (right) after 21-Gy single-dose irradiation. Very prominent TGF-β1 immunoreactivity and minimal TGF-β2 and TGF-β3 immunoreactivity is seen at both observation times.

Article Snippet: 31 Antibodies to pan-TGF-β (AB-100-NA), TGF-β2 (AB-12-NA), and TGF-β3 (AB-244-NA) were purchased from R&D Systems (Minneapolis, MN).

Techniques: Irradiation

Journal:

Article Title: Cellular Sources of Transforming Growth Factor-? Isoforms in Early and Chronic Radiation Enteropathy

doi:

Figure Lengend Snippet: In situ hybridization demonstrating TGF-β1 mRNA expression in irradiated intestine. A: TGF-β1 expression in regenerating crypt at the edge of a radiation-induced mucosal ulcer 2 weeks after 21-Gy single-dose irradiation. Magnification, ×200; bar, 100 μm. B: Inflammatory cells and fibroblast-like cells in the base of a radiation-induced mucosal ulcer expressing TGF-β1 2 weeks after 21-Gy single-dose irradiation. Magnification, ×400; bar, 50 μm. C: Intestinal wall beneath a radiation-induced mucosal ulcer (right overview; magnification, ×100; bar, 100 μm). Strong TGF-β1 expression in smooth muscle cells (left upper panel; magnification, ×400) and peritoneal mesothelium (left lower panel; magnification, ×400) is seen 2 weeks after 21-Gy single-dose irradiation. D: TGF-β1 expression in vascular endothelial cells and perivascular cells 26 weeks after 21-Gy single-dose irradiation. Magnification, ×400; bar, 50 μm. E: Fibroblasts in fibrotic area expressing TGF-β1 mRNA 26 weeks after 21-Gy single-dose irradiation. Magnification, ×400; bar, 50 μm.

Article Snippet: 31 Antibodies to pan-TGF-β (AB-100-NA), TGF-β2 (AB-12-NA), and TGF-β3 (AB-244-NA) were purchased from R&D Systems (Minneapolis, MN).

Techniques: In Situ Hybridization, Expressing, Irradiation

Journal:

Article Title: Cellular Sources of Transforming Growth Factor-? Isoforms in Early and Chronic Radiation Enteropathy

doi:

Figure Lengend Snippet: Expression of TGF-β mRNA in Sham-Irradiated and Irradiated Intestine, 2 and 26 Weeks after Irradiation

Article Snippet: 31 Antibodies to pan-TGF-β (AB-100-NA), TGF-β2 (AB-12-NA), and TGF-β3 (AB-244-NA) were purchased from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Irradiation

Journal: The Journal of Neuroscience

Article Title: Presynaptic Neuronal Nicotinic Receptors Differentially Shape Select Inputs to Auditory Thalamus and Are Negatively Impacted by Aging

doi: 10.1523/JNEUROSCI.1795-17.2017

Figure Lengend Snippet: nAChR β2 antibody labeling in WT and β2 KO mouse model. Immunofluorescence data in cortex of WT and β2 KO mouse model. Polyclonal rabbit anti-AChR β2 antibody (1:250, AB15325; Millipore) was used to label β2-containing nAChRs. WT shows greater β2 staining compared with KO. Remaining β2 immunolabel in KO is confined to nuclei. Negative controls in both show no β2 staining. Scale bar, 10 μm.

Article Snippet: Sections were transferred to primary antibody solution containing monoclonal mouse anti-vesicular glutamate transporter 1 (VGlut1) antibody (1:750; Millipore) and rabbit anti-nAChR β2 antibody (1:250; Millipore) and incubated overnight at room temperature.

Techniques: Antibody Labeling, Immunofluorescence, Staining, Immunolabeling

Journal: The Journal of Neuroscience

Article Title: Presynaptic Neuronal Nicotinic Receptors Differentially Shape Select Inputs to Auditory Thalamus and Are Negatively Impacted by Aging

doi: 10.1523/JNEUROSCI.1795-17.2017

Figure Lengend Snippet: Details of riboprobes targeted by FISH

Article Snippet: Sections were transferred to primary antibody solution containing monoclonal mouse anti-vesicular glutamate transporter 1 (VGlut1) antibody (1:750; Millipore) and rabbit anti-nAChR β2 antibody (1:250; Millipore) and incubated overnight at room temperature.

Techniques:

Journal: The Journal of Neuroscience

Article Title: Presynaptic Neuronal Nicotinic Receptors Differentially Shape Select Inputs to Auditory Thalamus and Are Negatively Impacted by Aging

doi: 10.1523/JNEUROSCI.1795-17.2017

Figure Lengend Snippet: nAChR β2 subunit colocalizes with VGluT1-positive corticothalamic terminals. Shown are representative confocal images of a distal dendrite from a Neurobiotin-labeled MGB neuron (cyan) from an excitatory neuron (VgluT1, green). The nAChR β2 subunit (red) labeled corticothalamic terminals apposing the MGB dendrite from a recorded neuron. Scale bar, 5 μm.

Article Snippet: Sections were transferred to primary antibody solution containing monoclonal mouse anti-vesicular glutamate transporter 1 (VGlut1) antibody (1:750; Millipore) and rabbit anti-nAChR β2 antibody (1:250; Millipore) and incubated overnight at room temperature.

Techniques: Labeling

Journal: The Journal of Neuroscience

Article Title: Presynaptic Neuronal Nicotinic Receptors Differentially Shape Select Inputs to Auditory Thalamus and Are Negatively Impacted by Aging

doi: 10.1523/JNEUROSCI.1795-17.2017

Figure Lengend Snippet: AC VGluT1-positive glutamatergic neurons coexpress β2 and α4 nAChR subunits. FISH in AC showing VGluT1 (blue), α4 nAChR subunit (red), β2 nAChR subunit (green), and DAPI (blue). A, Low-magnification image of infragranular projection layers LV, LVI, and white matter (wm). Scale bar, 100 μm. B, C, High-magnification images of VGluT1 projection neurons expressing α4 and β2 nAChR subunit mRNA (insets from A). Scale bar, 20 μm.

Article Snippet: Sections were transferred to primary antibody solution containing monoclonal mouse anti-vesicular glutamate transporter 1 (VGlut1) antibody (1:750; Millipore) and rabbit anti-nAChR β2 antibody (1:250; Millipore) and incubated overnight at room temperature.

Techniques: Expressing

Journal: The Journal of Neuroscience

Article Title: Presynaptic Neuronal Nicotinic Receptors Differentially Shape Select Inputs to Auditory Thalamus and Are Negatively Impacted by Aging

doi: 10.1523/JNEUROSCI.1795-17.2017

Figure Lengend Snippet: IC VGAT-positive GABAergic neurons express β2, but not α4, nAChR subunits, whereas some VGAT-negative cells coexpress both. FISH in the IC shows VGAT (blue), α4 nAChR subunit (red), β2 nAChR subunit (green), and DAPI (white). A, Low-magnification image montage of the IC. Scale bar, 250 μm. B, C, High-magnification images of insets from A showing blue VGAT-labeled large inhibitory, putative projection neurons expressing β2 nAChR subunit mRNA and unlabeled putative glutamatergic neurons expressing α4 and β2 nAChR subunit mRNAs. Scale bar, 20 μm.

Article Snippet: Sections were transferred to primary antibody solution containing monoclonal mouse anti-vesicular glutamate transporter 1 (VGlut1) antibody (1:750; Millipore) and rabbit anti-nAChR β2 antibody (1:250; Millipore) and incubated overnight at room temperature.

Techniques: Labeling, Expressing

Journal: The Journal of Neuroscience

Article Title: Presynaptic Neuronal Nicotinic Receptors Differentially Shape Select Inputs to Auditory Thalamus and Are Negatively Impacted by Aging

doi: 10.1523/JNEUROSCI.1795-17.2017

Figure Lengend Snippet: Major inputs to the MGB and their neurotransmitters. The responses of MGB neurons are dictated by excitatory and inhibitory sensory inputs from IC, excitatory input from AC, and inhibitory input from TRN. ACh released from PPTg may then activate nAChRs within the MGB circuitry. Inset shows proposed nAChR sites for cholinergic modulation/activation of MGB neurons. Shown are postsynaptic nAChRs on MGB neurons (Sottile et al., 2017) and presynaptic nAChRs on excitatory corticothalamic input, both of which are likely to be α4β2 nAChRs, whereas presynaptic nAChRs on inhibitory tectothalamic input are β2-containing nAChRs with a different α subunit.

Article Snippet: Sections were transferred to primary antibody solution containing monoclonal mouse anti-vesicular glutamate transporter 1 (VGlut1) antibody (1:750; Millipore) and rabbit anti-nAChR β2 antibody (1:250; Millipore) and incubated overnight at room temperature.

Techniques: Activation Assay

Journal: PeerJ

Article Title: CircHIPK3 regulates cardiac fibroblast proliferation, migration and phenotypic switching through the miR-152-3p/TGF-β2 axis under hypoxia

doi: 10.7717/peerj.9796

Figure Lengend Snippet: To determine the role of miR-152-3p in CFs, CFs were transfected with the miR-152-3p mimic or inhibitor. (A–G and J–P) Representative images of EdU-stained CFs from different groups. Quantification of EdU+ cells presented as the % EdU-positive cells and Hoechst-stained nuclei. The orange color is EdU-positive cells, and the blue color is nuclei stained by Hoechst 33342. Scale bar = 20 μm. n = 10. * P < 0.05, vs. MNC group or INC group. (H, I and Q, R) The protein expression levels of Col I and Col III were measured by western blot analysis. n = 3. * P < 0.05, vs. MNC group or INC group. (S) Relative expression of miR-152-3p in CFs after transfection with miR-152-3p mimic or inhibitor was examined by RT-qPCR analysis. n = 3. * P < 0.05, vs. hypoxia group. (T) TGF-β2 was predicted as a target gene of miR-152-3p using the TargetScan database. HEK-293T cells were cotransfected with RLuc-TGF-β2-WT or RLuc-TGF-β2-Mut and miR-152-3p mimics or NC mimic. Luciferase activity was detected using the dual luciferase reporter assay at 48 h post-transfection. n = 3. * P < 0.05, vs. TGF-β2-WT + NC group. # P < 0.05, vs. TGF-β2-WT + miR-152-3p group. (U–X) The protein expression level of TGF-β2 was measured by western blot analysis. n = 3. * P < 0.05, vs. MNC group or INC group.

Article Snippet: The primary anti-bodies: Anti-Collagen I (Cell Signaling Technology, 91144, dilution: 1:1,000), Anti-Collagen III (Abcam, ab7778, dilution: 1:7,000), Anti-α-SMA (Cell Signaling Technology, 19245, dilution: 1:1,000), Anti-p-Smad2 (Cell Signaling Technology, 18338, dilution: 1:1,000), Anti-p-Smad3 (Cell Signaling Technology, 9520, dilution: 1:1,000), Anti-TGF-β2 (Bioss, bs-20412R, dilution: 1:1,000) and Anti-β-actin Antibody (Abcam, ab8227, dilution: 1:1,000), were used.

Techniques: Transfection, Staining, Expressing, Western Blot, Quantitative RT-PCR, Luciferase, Activity Assay, Reporter Assay

Journal: PeerJ

Article Title: CircHIPK3 regulates cardiac fibroblast proliferation, migration and phenotypic switching through the miR-152-3p/TGF-β2 axis under hypoxia

doi: 10.7717/peerj.9796

Figure Lengend Snippet: (A–E) The cell cycle was investigated by a cell cycle assay in cardiac fibroblasts. The results represent the percentage of cells in G1 and S + G2/M phases for the indicated conditions. n = 3. * P < 0.05, # P < 0.05. (F–J) Wound scratch assay for the different groups. The average sizes of the gaps were measured at 48 h. Scale bar = 200 μm. n = 5. * P < 0.05, # P < 0.05. (K and L) The protein expression levels of Col I, Col III, TGF-β2, p-Smad2 and p-Smad3 were measured by western blot analysis. n = 3. * P < 0.05, # P < 0.05. * P < 0.05, vs. hypoxia group, # P < 0.05, vs. si-circHIPK3 group.

Article Snippet: The primary anti-bodies: Anti-Collagen I (Cell Signaling Technology, 91144, dilution: 1:1,000), Anti-Collagen III (Abcam, ab7778, dilution: 1:7,000), Anti-α-SMA (Cell Signaling Technology, 19245, dilution: 1:1,000), Anti-p-Smad2 (Cell Signaling Technology, 18338, dilution: 1:1,000), Anti-p-Smad3 (Cell Signaling Technology, 9520, dilution: 1:1,000), Anti-TGF-β2 (Bioss, bs-20412R, dilution: 1:1,000) and Anti-β-actin Antibody (Abcam, ab8227, dilution: 1:1,000), were used.

Techniques: Cell Cycle Assay, Wound Healing Assay, Expressing, Western Blot