Journal: Cell Biology and Toxicology

Article Title: TRAF6 promotes spinal microglial M1 polarization to aggravate neuropathic pain by activating the c-JUN/NF-kB signaling pathway

doi: 10.1007/s10565-024-09900-6

Figure Lengend Snippet: Inhibition of TRAF6 relieved hyperalgesia induced by CCI. ( A ) The western blot and qRT-PCR assay tested AAV-shRNA or AAV-shNC transfection efficiency in CCI-induced mice. ( B-D ) The immunofluorescence was further employed to detect co-expression of TRAF6 with NeuN, GFAP, and Iba-1, respectively, in CCI-induced mice after intrathecally injecting AAV-shRNA or AAV-shNC. ( E ) Mechanical allodynia was estimated by Von Frey filaments detection of paw withdrawal thresholds, and thermal hyperalgesia was calculated by radiant heat detection of paw withdrawal latency. ** p < 0.01 vs Sham, and # p < 0.05, ## p < 0.01 vs CCI. Scale bar: 200/100 μm

Article Snippet: Fixed spinal cord tissues were paraffin-embedded, deparaffinized and repaired with antigens (boiled, 10 min), blocked in goat serum (room temperature, Solarbio,15 min), and incubated with primary antibodies (4 °C, 24 h): anti-TRAF6 (A0973, ABclonal, 1:1000), anti-iNOS (Abcam, ab178945, 1:500), anti-Arg1 (Abcam, ab96183, 1:500), anti-Iba-1 (Abcam, ab178846, 1:500), Neuronal Marker anti-NeuN (Abcam, ab177487, 1:1000), anti-GFAP (Proteintech, 16825–1-AP, 1:50).

Techniques: Inhibition, Western Blot, Quantitative RT-PCR, shRNA, Transfection, Immunofluorescence, Expressing

Journal: bioRxiv

Article Title: Cerebellar contribution to cognitive deficits and prefrontal cortex dysfunction in Spinocerebellar Ataxia Type 1 (SCA1)

doi: 10.1101/2024.07.10.602931

Figure Lengend Snippet: Saggital brain sections of 26 week old ATXN1[82] mice and their wild-type littermates were immunostained for NeuN, c-Fos, VGLUT2, PSD95, VGAT and gephyrin, and confocal z-stack images were obtained. A. Quantification of NeuN density. B. Quantification of c-Fos density. C. Image of VGLUT2 and PSD95 puncta. D. Quantification of excitatory presynaptic VGLUT2+, postsynaptic PSD95+ and co-localized VGLUT2/PSD95 puncta. E. Quantification of inhibitory presynaptic VGAT+, postsynaptic Gephyrin+ and co-localized VGAT/gephyrin puncta. N= 6-12 ATXN1[82] mice and their wild-type littermates. * P< 0.05, **** P<0.0001 Student’s t-test.

Article Snippet: We used primary antibodies against c-Fos (rabbit, Abcam, ab190289), neuronal marker neuronal nuclei (NeuN) (rabbit, Abcam, Ab104225), PSD95 (rabbit, Thermo Fisher Scientific, 51-69000), vesicular glutamate transporter 2 (VGLUT2) (guinea pig, Millipore, AB2251-I), Gephyrin (rabbit, Thermo Fisher Scientific, PA5-29036), and VGAT (guinea pig, Synaptic Systems,131005) as previously described , .

Techniques:

Journal: bioRxiv

Article Title: Cerebellar contribution to cognitive deficits and prefrontal cortex dysfunction in Spinocerebellar Ataxia Type 1 (SCA1)

doi: 10.1101/2024.07.10.602931

Figure Lengend Snippet: Saggital brain sections of 18-week-old f-ATXN1 146Q mice and their wild-type littermates were immunostained for NeuN, c-Fos, VGLUT2, PSD95, VGAT and gephyrin, and confocal z-stack images were obtained. A. Quantification of NeuN density. B. Quantification of c-Fos density. C. Image of VGLUT2 and PSD95 puncta. D. Quantification of excitatory presynaptic VGLUT2+, postsynaptic PSD95+ and co-localized VGLUT2/PSD95 puncta. E. Quantification of inhibitory presynaptic VGAT+, postsynaptic Gephyrin+ and co-localized VGAT/gephyrin puncta. N= 6-12 f-ATXN1 146Q mice and their wild-type littermates. * P< 0.05, **** P<0.0001 Student’s t-test.

Article Snippet: We used primary antibodies against c-Fos (rabbit, Abcam, ab190289), neuronal marker neuronal nuclei (NeuN) (rabbit, Abcam, Ab104225), PSD95 (rabbit, Thermo Fisher Scientific, 51-69000), vesicular glutamate transporter 2 (VGLUT2) (guinea pig, Millipore, AB2251-I), Gephyrin (rabbit, Thermo Fisher Scientific, PA5-29036), and VGAT (guinea pig, Synaptic Systems,131005) as previously described , .

Techniques: