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CPTP induced upregulation of <t>astroglial</t> <t>C3</t> in the spinal dorsal horn (A) Mechanical withdrawal threshold (MWT) was measured in the ipsilateral (CPTP-ipsi) and contralateral (CPTP-contra) sides of rats compared with sham-operated controls (Sham-ipsi and Sham-contra). n = 6, two-way ANOVA following Bonferroni’s post hoc test, ∗∗∗∗ p < 0.0001, vs. CPTP-contra; ## p < 0.01, ### p < 0.001, #### p < 0.0001, vs. Sham-ipsi. (B) Measurements of hind paw scratches and turning maneuvers challenged by cold acetone. n = 6, two-way ANOVA following Bonferroni’s post hoc test, ∗∗∗∗ p < 0.0001, vs. CPTP-contra; #### p < 0.0001, vs. Sham-ipsi. (C) <t>ELISA</t> analysis of C3 concentration in the CSF from sham and CPTP rats on POD 14. n = 4, Student’s t test, ∗∗ p < 0.01. (D) qRT-PCR analysis of C3 in the TDH from sham and CPTP rats on POD 14. n = 4, Student’s t test, ∗ p < 0.05. (E) Western blot analysis of C3 in the TDH from naive, sham, and CPTP rats on POD 7, 14, and 21. n = 4, One-way ANOVA following Bonferroni’s post hoc test, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, vs. Sham; # p < 0.05, ## p < 0.01, ### p < 0.001, vs. Naive. (F and G) Representative images of TDH on POD 14 stained for C3 (green), GFAP (red), along with the quantitative analysis of percentage overlap. n = 3, Student’s t test, ∗∗∗∗ p < 0.0001. Scale bars: 50 μm. See also and for the schematic of the CPTP model and C3 + cell distribution in the TDH. Data are presented as mean ± SEM.
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CPTP induced upregulation of astroglial C3 in the spinal dorsal horn (A) Mechanical withdrawal threshold (MWT) was measured in the ipsilateral (CPTP-ipsi) and contralateral (CPTP-contra) sides of rats compared with sham-operated controls (Sham-ipsi and Sham-contra). n = 6, two-way ANOVA following Bonferroni’s post hoc test, ∗∗∗∗ p < 0.0001, vs. CPTP-contra; ## p < 0.01, ### p < 0.001, #### p < 0.0001, vs. Sham-ipsi. (B) Measurements of hind paw scratches and turning maneuvers challenged by cold acetone. n = 6, two-way ANOVA following Bonferroni’s post hoc test, ∗∗∗∗ p < 0.0001, vs. CPTP-contra; #### p < 0.0001, vs. Sham-ipsi. (C) ELISA analysis of C3 concentration in the CSF from sham and CPTP rats on POD 14. n = 4, Student’s t test, ∗∗ p < 0.01. (D) qRT-PCR analysis of C3 in the TDH from sham and CPTP rats on POD 14. n = 4, Student’s t test, ∗ p < 0.05. (E) Western blot analysis of C3 in the TDH from naive, sham, and CPTP rats on POD 7, 14, and 21. n = 4, One-way ANOVA following Bonferroni’s post hoc test, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, vs. Sham; # p < 0.05, ## p < 0.01, ### p < 0.001, vs. Naive. (F and G) Representative images of TDH on POD 14 stained for C3 (green), GFAP (red), along with the quantitative analysis of percentage overlap. n = 3, Student’s t test, ∗∗∗∗ p < 0.0001. Scale bars: 50 μm. See also and for the schematic of the CPTP model and C3 + cell distribution in the TDH. Data are presented as mean ± SEM.

Journal: iScience

Article Title: Astrocyte-to-neuron interaction via NF-κB/C3/C3aR mediates chronic post-thoracotomy pain by modulating neuronal GluR1 in spinal dorsal horn

doi: 10.1016/j.isci.2025.113917

Figure Lengend Snippet: CPTP induced upregulation of astroglial C3 in the spinal dorsal horn (A) Mechanical withdrawal threshold (MWT) was measured in the ipsilateral (CPTP-ipsi) and contralateral (CPTP-contra) sides of rats compared with sham-operated controls (Sham-ipsi and Sham-contra). n = 6, two-way ANOVA following Bonferroni’s post hoc test, ∗∗∗∗ p < 0.0001, vs. CPTP-contra; ## p < 0.01, ### p < 0.001, #### p < 0.0001, vs. Sham-ipsi. (B) Measurements of hind paw scratches and turning maneuvers challenged by cold acetone. n = 6, two-way ANOVA following Bonferroni’s post hoc test, ∗∗∗∗ p < 0.0001, vs. CPTP-contra; #### p < 0.0001, vs. Sham-ipsi. (C) ELISA analysis of C3 concentration in the CSF from sham and CPTP rats on POD 14. n = 4, Student’s t test, ∗∗ p < 0.01. (D) qRT-PCR analysis of C3 in the TDH from sham and CPTP rats on POD 14. n = 4, Student’s t test, ∗ p < 0.05. (E) Western blot analysis of C3 in the TDH from naive, sham, and CPTP rats on POD 7, 14, and 21. n = 4, One-way ANOVA following Bonferroni’s post hoc test, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, vs. Sham; # p < 0.05, ## p < 0.01, ### p < 0.001, vs. Naive. (F and G) Representative images of TDH on POD 14 stained for C3 (green), GFAP (red), along with the quantitative analysis of percentage overlap. n = 3, Student’s t test, ∗∗∗∗ p < 0.0001. Scale bars: 50 μm. See also and for the schematic of the CPTP model and C3 + cell distribution in the TDH. Data are presented as mean ± SEM.

Article Snippet: Samples were centrifuged at 5000 g for 3 min at 4°C, and supernatants were analyzed for C3 levels using an ELISA kit (Elabscience, E-EL-R0250).

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Quantitative RT-PCR, Western Blot, Staining