Journal: STAR Protocols

Article Title: Protocol for handling and using SOD1 mice for amyotrophic lateral sclerosis pre-clinical studies

doi: 10.1016/j.xpro.2024.103459

Figure Lengend Snippet:

Article Snippet: Rabbit anti-Chat (Genetex) , Staining of motor neurons , 1:750.

Techniques: Staining

Journal: Neurobiology of Stress

Article Title: The insular cortex-nucleus tractus solitarius glutamatergic pathway involved in acute stress-induced gastric mucosal damage in rats

doi: 10.1016/j.ynstr.2025.100723

Figure Lengend Snippet: Inhibition IC glutamate neurons to the influence of the gastric mucosa damage induced by RWIS. A: Schematic of virus injection; B–C: Viral expression in the left and right IC (40 × ) scale bar, 200 μm; D–E: Viral expression in the left and right IC (100 × ) scale bar, 100 μm; F: c-Fos expression in RWIS rats treated with CaMKIIa-mCherry in the IC; G–H: c-Fos expression in RWIS rats in the IC of the CaMKIIa-hm4d + saline- or CNO-injected groups (100 × ). scale bar,100 μm, c-Fos in a rectangle were magnified to a higher magnification (400 × ); scale bar, 20 μm; I: The number of c-Fos-IR neurons in the IC was quantified. compared with mCherry group, ∗: P < 0.05, ∗∗: P < 0.01, compared with hm4d + SA group, #: P < 0.05, ##: P < 0.01 (n = 6); J: Gastric mucosal damage in RWIS rats treated with CaMKIIa-mCherry; K–L: Gastric mucosal damage in RWIS rats in the IC of the CaMKIIa-hm4d + saline- or CNO-injected groups; M: Quantification of erosion index (EI). compared with mCherry group, ∗: P < 0.05, ∗∗: P < 0.0, compared with hm4d + SA group, #: P < 0.05, ##: P < 0.01 (n = 6); N: Western blot of Occludin, Claudin-1, PCNA, NOS and CHAT protein expression in different groups; O–S: Quantification of the relative average grayscale value of Occludin/β-actin, Claudin-1/β-actin, PCNA/β-actin, NOS/β-actin and CHAT/β-actin. compared with the mCherry group, ∗: P < 0.05, ∗∗: P < 0.01, compared with hm4d + SA group, #: P < 0.05, ##: P < 0.01 (n = 5).

Article Snippet: After incubation at 4 °C overnight, rabbit anti-β-actin (1:8000; Huabio; Hangzhou, China), rabbit Occludin (1:1000; Huabio; Hangzhou, China), mouse anti- Claudin-1 (1:1000; Huabio; Hangzhou, China), rabbit anti-PCNA(1:5000; Proteintech,USA), rabbit anti-nNOS(1:500; Huabio; Hangzhou, China), rabbit anti-CHAT(1:500; Huabio; Hangzhou, China), rabbit anti-PSD95 (1:1000; Huabio; Hangzhou, China), rabbit anti-SYN-1 (1:1000; Huabio; Hangzhou, China).

Techniques: Inhibition, Virus, Injection, Expressing, Saline, Western Blot

Journal: Neurobiology of Stress

Article Title: The insular cortex-nucleus tractus solitarius glutamatergic pathway involved in acute stress-induced gastric mucosal damage in rats

doi: 10.1016/j.ynstr.2025.100723

Figure Lengend Snippet: Activation IC glutamate neurons to the influence of the gastric mucosa damage induced by RWIS. A: Schematic of virus injection; B–C Viral expression in the left and right IC (40 × ) scale bar, 200 μm; D–E: Viral expression in the left and right IC (100 × ) scale bar,100 μm; F: c-Fos expression in RWIS rats treated with CaMKIIa-mCherry in the IC; G–H: c-Fos expression in RWIS rats in the IC of the CaMKIIa-hm3d + saline- or CNO-injected groups (100 × ) scale bar,100 μm, c-Fos in a rectangle were magnified to a higher magnification (400 × ); scale bar, 20 μm; I: The number of c-Fos-IR neurons in the IC was quantified. compared with mCherry group, ∗: P < 0.05, ∗∗: P < 0.01, compared with hm3d + SA group, #: P < 0.05, ##: P < 0.01 (n = 6); J: Gastric mucosal damage in RWIS rats treated with CaMKIIa-mCherry; K–L: Gastric mucosal damage in RWIS rats in the IC of the CaMKIIa-hm3d + saline- or CNO-injected groups; M: Quantification of erosion index (EI). compared with mCherry group, ∗: P < 0.05, ∗∗: P < 0.0, compared with hm3d + SA group, #: P < 0.05, ##: P < 0.01 (n = 6); N: Western blot of Occludin, Claudin-1, PCNA, NOS and CHAT protein expression in different groups; O–S: Quantification of the relative average grayscale value of Occludin/β-actin, Claudin-1/β-actin, PCNA/β-actin, NOS/β-actin and CHAT/β-actin. compared with the mCherry group, ∗: P < 0.05, ∗∗: P < 0.01, compared with hm3d + SA group, #: P < 0.05, ##: P < 0.01 (n = 6).

Article Snippet: After incubation at 4 °C overnight, rabbit anti-β-actin (1:8000; Huabio; Hangzhou, China), rabbit Occludin (1:1000; Huabio; Hangzhou, China), mouse anti- Claudin-1 (1:1000; Huabio; Hangzhou, China), rabbit anti-PCNA(1:5000; Proteintech,USA), rabbit anti-nNOS(1:500; Huabio; Hangzhou, China), rabbit anti-CHAT(1:500; Huabio; Hangzhou, China), rabbit anti-PSD95 (1:1000; Huabio; Hangzhou, China), rabbit anti-SYN-1 (1:1000; Huabio; Hangzhou, China).

Techniques: Activation Assay, Virus, Injection, Expressing, Saline, Western Blot

Journal: Neurobiology of Stress

Article Title: The insular cortex-nucleus tractus solitarius glutamatergic pathway involved in acute stress-induced gastric mucosal damage in rats

doi: 10.1016/j.ynstr.2025.100723

Figure Lengend Snippet: Effect of inhibition of the IC-NTS glutamatergic pathway on RWIS-induced gastric mucosal damage. A: Schematic of virus injection; B: Viral expression in the IC (40 × ) scale bar, 200 μm; C: Viral expression in the IC (100 × ) scale bar,100 μm; D: c-Fos expression in RWIS rats treated with CaMKIIa-mCherry in the IC; E–F: c-Fos expression in RWIS rats in the IC of the CaMKIIa-hm4d + saline- or CNO-injected groups (100 × ). scale bar,100 μm, c-Fos in a rectangle were magnified to a higher magnification (400 × ); scale bar, 20 μm; G: The number of c-Fos-IR neurons in the insul was quantified. compared with mCherry group, ∗: P < 0.05, ∗∗: P < 0.01, compared with hm4d + SA group, #: P < 0.05, ##: P < 0.01 (n = 6); H: c-Fos expression in RWIS rats treated with CaMKIIa-mCherry in the NTS; I–J: c-Fos expression in RWIS rats in the NTS of the CaMKIIa-hm4d + saline- or CNO-injected groups (100 × ). scale bar,100 μm; K: The number of c-Fos-IR neurons in the NTS was quantified. compared with mCherry group, ∗: P < 0.05, ∗∗: P < 0.01, compared with hm4d + SA group, #: P < 0.05, ##: P < 0.01 (n = 6); L: Gastric mucosal damage in RWIS rats treated with CaMKIIa-mCherry; M–N: Gastric mucosal damage in RWIS rats in the IC of the CaMKIIa-hm4d + saline- or CNO-injected groups; O: Quantification of erosion index (EI). compared with mCherry group, ∗: P < 0.05, ∗∗: P < 0.0, compared with hm4d + SA group, #: P < 0.05, ##: P < 0.01 (n = 6); P: Western blot of Occludin, Claudin-1, PCNA, NOS and CHAT protein expression in different groups; Q–U: Quantification of the relative average grayscale value of Occludin/β-actin, Claudin-1/β-actin, PCNA/β-actin, NOS/β-actin and CHAT/β-actin. compared with the mCherry group, ∗: P < 0.05, ∗∗: P < 0.01, compared with hm4d + SA group, #: P < 0.05, ##: P < 0.01 (n = 6).

Article Snippet: After incubation at 4 °C overnight, rabbit anti-β-actin (1:8000; Huabio; Hangzhou, China), rabbit Occludin (1:1000; Huabio; Hangzhou, China), mouse anti- Claudin-1 (1:1000; Huabio; Hangzhou, China), rabbit anti-PCNA(1:5000; Proteintech,USA), rabbit anti-nNOS(1:500; Huabio; Hangzhou, China), rabbit anti-CHAT(1:500; Huabio; Hangzhou, China), rabbit anti-PSD95 (1:1000; Huabio; Hangzhou, China), rabbit anti-SYN-1 (1:1000; Huabio; Hangzhou, China).

Techniques: Inhibition, Virus, Injection, Expressing, Saline, Western Blot

Journal: Neurobiology of Stress

Article Title: The insular cortex-nucleus tractus solitarius glutamatergic pathway involved in acute stress-induced gastric mucosal damage in rats

doi: 10.1016/j.ynstr.2025.100723

Figure Lengend Snippet: Effect of activation of the IC-NTS glutamatergic pathway on RWIS-induced gastric mucosal damage. A: Schematic of virus injection; B: viral expression in the IC (40 × ) scale bar, 200 μm; C: viral expression in the IC (100 × ) scale bar,100 μm; D: c-Fos expression in RWIS rats treated with CaMKIIa-mCherry in the IC; E–F: c-Fos expression in RWIS rats in the IC of the CaMKIIa-hm3d + saline- or CNO-injected groups (100 × ). scale bar,100 μm, c-Fos in a rectangle were magnified to a higher magnification (400 × ); scale bar, 20 μm; G: The number of c-Fos-IR neurons in the insul was quantified. compared with mCherry group, ∗: P < 0.05, ∗∗: P < 0.01, compared with hm3d + SA group, #: P < 0.05, ##: P < 0.01 (n = 6); H: c-Fos expression in RWIS rats treated with CaMKIIa-mCherry in the NTS; I–J: c-Fos expression in RWIS rats in the NTS of the CaMKIIa-hm3d + saline- or CNO-injected groups (100 × ). scale bar,100 μm; K: The number of c-Fos-IR neurons in the NTS was quantified. compared with mCherry group, ∗: P < 0.05, ∗∗: P < 0.01, compared with hm3d + SA group, #: P < 0.05, ##: P < 0.01 (n = 6); L: Gastric mucosal damage in RWIS rats treated with CaMKIIa-mCherry; M–N: Gastric mucosal damage in RWIS rats in the IC of the CaMKIIa-hm3d + saline- or CNO-injected groups; O: Quantification of erosion index (EI). compared with mCherry group, ∗: P < 0.05, ∗∗: P < 0.0, compared with hm3d + SA group, #: P < 0.05, ##: P < 0.01 (n = 6); P: Western blot of Occludin, Claudin-1, PCNA, NOS and CHAT protein expression in different groups; Q–U: Quantification of the relative average grayscale value of Occludin/β-actin, Claudin-1/β-actin, PCNA/β-actin, NOS/β-actin and CHAT/β-actin. compared with the mCherry group, ∗: P < 0.05, ∗∗: P < 0.01, compared with hm3d + SA group, #: P < 0.05, ##: P < 0.01 (n = 6).

Article Snippet: After incubation at 4 °C overnight, rabbit anti-β-actin (1:8000; Huabio; Hangzhou, China), rabbit Occludin (1:1000; Huabio; Hangzhou, China), mouse anti- Claudin-1 (1:1000; Huabio; Hangzhou, China), rabbit anti-PCNA(1:5000; Proteintech,USA), rabbit anti-nNOS(1:500; Huabio; Hangzhou, China), rabbit anti-CHAT(1:500; Huabio; Hangzhou, China), rabbit anti-PSD95 (1:1000; Huabio; Hangzhou, China), rabbit anti-SYN-1 (1:1000; Huabio; Hangzhou, China).

Techniques: Activation Assay, Virus, Injection, Expressing, Saline, Western Blot

Journal: Regenerative Therapy

Article Title: Enhanced axon outgrowth of spinal motor neurons in co-culturing with dorsal root ganglions antagonizes the growth inhibitory environment

doi: 10.1016/j.reth.2023.11.013

Figure Lengend Snippet: A: The SMNs in dispersed culture on the 5th day (upper). 3D printed mold and PDMS template for SMNA preparation (lower left) and the RFP-transduced SMNA plated on the 3rd day (lower right). B: The cultured SMNs were immunofluorescently identified as positive for ChAT, Tuj1 and NeuN. C: Different degrees of axonal connections shown at two populations of SMNAs plated at various distances. Robust axonal connections formed in 2 mm of distance (upper), a small number of axonal connections formed in 3 mm of distance (middle) and no axonal connection was observed in 4 mm of distance (lower). Scale Bar: A, B: 100 μm C: 200 μm.

Article Snippet: Then, the rabbit anti-ChAT antibody (Beyotime, AF6497, 1:500), mouse anti-neuron-specific Class III β-tubulin antibody (Tuj1, Covance, MMS-435P, 1:500), or rabbit anti-NeuN (Proteintech, 26975-1-AP, 1:500) was added to the cell slides for incubation overnight at 4 °C, followed by incubation for 30 min at 37 °C with Cy3-labelled goat anti-rabbit IgG (H & L) (Beyotime, A0516, 1:500), Cy3-labelled goat anti-mouse IgG (H & L) (Beyotime, A0521, 1:500) or Dylight 488-labelled Goat anti-rabbit IgG (H & L) (Abcam, ab96883, 1:1000) as the second antibody, respectively.

Techniques: Cell Culture