Journal: International Journal of Molecular Sciences

Article Title: Pannexin 1-Mediated ATP Signaling in the Trigeminal Spinal Subnucleus Caudalis Is Involved in Tongue Cancer Pain

doi: 10.3390/ijms222111404

Figure Lengend Snippet: The possible involvement of PANX1 in the Vc in mechanical allodynia in the tongue following SCC inoculation. ( A ) Time course of mechanical head-withdrawal threshold with stimulation of the tongue in PBS-injected or SCC-inoculated rats. n = 8: PBS, n = 12: SCC, ** p < 0.01, *** p < 0.001, vs. Pre, one-way ANOVA post hoc Dunnet’s test; †† p < 0.01, ††† p < 0.001, vs. PBS, two-way repeated-measures ANOVA followed by Bonferroni’s test. Data represent the mean ± SEM. ( B ) Time course of mechanical head-withdrawal threshold with stimulation of the tongue in PBS-injected or SCC-inoculated rats with intracisternal administration of PBS or 10Panx. n = 9: PBS + PBS, n = 6: SCC + PBS, n = 10: SCC + 10 Panx, ** p < 0.01, *** p < 0.001, vs. SCC + PBS, two-way repeated-measures ANOVA followed by Bonferroni’s test. Data represent the mean ± SEM.

Article Snippet: The slices were incubated with rabbit polyclonal anti-PANX1 antibody (1:50; Cat. MBS2001670, MYBioSource, San Diego, CA, USA), goat polyclonal anti-Iba1 antibody (1:1000; Cat. ab5076, Abcam, Cambridge, MA, USA), mouse monoclonal anti-GFAP antibody (1:1000; Cat. MAB360, Merck Millipore, Billerica, MA, USA), or mouse monoclonal anti-NeuN antibody (1:100; Cat. MAB377; Merck Millipore) for 2 days at 4 °C.

Techniques: Injection

Journal: International Journal of Molecular Sciences

Article Title: Pannexin 1-Mediated ATP Signaling in the Trigeminal Spinal Subnucleus Caudalis Is Involved in Tongue Cancer Pain

doi: 10.3390/ijms222111404

Figure Lengend Snippet: Expression of PANX1 in the Vc. ( A , B ) PANX-IR cells in the Vc 14 days after PBS injection with intracisternal (i.c.) administration of PBS (PBS + PBS) or SCC inoculation with PBS (i.c.) (SCC + PBS). Scale bar = 100 μm. ( C ) Columns represent the average values of area occupied by PANX1 immuno-product. n = 5: PBS + PBS, n = 5: SCC + PBS, unpaired Student’s t -test, ** p < 0.01. Data represent the mean ± SEM. ( D – F ) Representative images showing PANX1 (green) and Iba1, GFAP, or NeuN (red) in the Vc. Arrowheads indicate PANX1/Iba1 or PANX1/GFAP double-positive cells. Scale bar = 100 μm. ( G ) Representative PANX1 image in the Vc. Insets were set in the lamina I-II (a), III-IV (b) and V (c). Scale bar = 500 μm. ( H ) Columns represent the average number of PANX1/Iba1 double-positive cells in each lamina. a–c correspond to the inset in ( G ). n = 5: PBS + PBS (i.c.), n = 5: SCC + PBS (i.c.), unpaired Student’s t -test, ** p < 0.01. Data represent the mean ± SEM.

Article Snippet: The slices were incubated with rabbit polyclonal anti-PANX1 antibody (1:50; Cat. MBS2001670, MYBioSource, San Diego, CA, USA), goat polyclonal anti-Iba1 antibody (1:1000; Cat. ab5076, Abcam, Cambridge, MA, USA), mouse monoclonal anti-GFAP antibody (1:1000; Cat. MAB360, Merck Millipore, Billerica, MA, USA), or mouse monoclonal anti-NeuN antibody (1:100; Cat. MAB377; Merck Millipore) for 2 days at 4 °C.

Techniques: Expressing, Injection

Journal: International Journal of Molecular Sciences

Article Title: Pannexin 1-Mediated ATP Signaling in the Trigeminal Spinal Subnucleus Caudalis Is Involved in Tongue Cancer Pain

doi: 10.3390/ijms222111404

Figure Lengend Snippet: PANX1 is required for the enhancement of firing activity in Vc neurons following SCC inoculation. ( A ) Representative image showing the brainstem. An illustration showing the transverse plane of the brainstem. The black circles indicate the area that responds to the tongue stimulation. ( B ) The RFs of the tongue 14 days after PBS injection with intracisternal (i.c.) administration of PBS (PBS + PBS), SCC inoculation with PBS (i.c.) (SCC + PBS), or SCC inoculation with 10Panx (i.c.) (SCC + 10Panx). The area surrounded by the red lines indicates the area that responds to the stimulus. Columns represent the average ratio of RF in the tongue. ( C ) Representative firing activities following stimulation with brush, non-noxious, noxious and pinch stimulation in PBS + PBS, SCC + PBS and SCC + 10Panx rats. ( D – I ) Columns represent the average number of spikes of background activity ( D ) and afterdischarge ( E ) and brush ( F ), non-noxious pressure ( G ), noxious pressure ( H ) and pinch ( I ) induced spikes in PBS + PBS, SCC + PBS and SCC + 10Panx rats. n = 5: PBS + PBS, n = 6: SCC + PBS, n = 4: SCC + 10Panx, * p < 0.05, one-way ANOVA followed by Tukey’s test. Data represent the mean ± SEM.

Article Snippet: The slices were incubated with rabbit polyclonal anti-PANX1 antibody (1:50; Cat. MBS2001670, MYBioSource, San Diego, CA, USA), goat polyclonal anti-Iba1 antibody (1:1000; Cat. ab5076, Abcam, Cambridge, MA, USA), mouse monoclonal anti-GFAP antibody (1:1000; Cat. MAB360, Merck Millipore, Billerica, MA, USA), or mouse monoclonal anti-NeuN antibody (1:100; Cat. MAB377; Merck Millipore) for 2 days at 4 °C.

Techniques: Activity Assay, Injection

Journal: International Journal of Molecular Sciences

Article Title: Pannexin 1-Mediated ATP Signaling in the Trigeminal Spinal Subnucleus Caudalis Is Involved in Tongue Cancer Pain

doi: 10.3390/ijms222111404

Figure Lengend Snippet: PANX1 contributes to the increment of phosphorylation of ERK in Vc neurons following SCC inoculation. ( A – C ) The representative images showing pERK Vc neurons in PBS + PBS, SCC + PBS and SCC + 10Panx rats on day 14 after inoculation. Scale bar = 200 μm. ( D ) Columns represent the average number of pERK-IR neurons in the Vc. Arrowheads indicate pERK-IR neurons. n = 5: PBS + PBS, n = 5: SCC + PBS, n = 5: SCC + 10Panx, * p < 0.05, one-way ANOVA followed by Tukey’s test. Data represent the mean ± SEM.

Article Snippet: The slices were incubated with rabbit polyclonal anti-PANX1 antibody (1:50; Cat. MBS2001670, MYBioSource, San Diego, CA, USA), goat polyclonal anti-Iba1 antibody (1:1000; Cat. ab5076, Abcam, Cambridge, MA, USA), mouse monoclonal anti-GFAP antibody (1:1000; Cat. MAB360, Merck Millipore, Billerica, MA, USA), or mouse monoclonal anti-NeuN antibody (1:100; Cat. MAB377; Merck Millipore) for 2 days at 4 °C.

Techniques: Phospho-proteomics