Journal: eBioMedicine

Article Title: Reprogramming astrocytic NDRG2/NF-κB/C3 signaling restores the diabetes-associated cognitive dysfunction

doi: 10.1016/j.ebiom.2023.104653

Figure Lengend Snippet: Excluded variables of multivariate logistic regression analyses.

Article Snippet: The Human C3 (Complement Component 3) ELISA Kit (E-EL-H6054) and Human INS (Insulin) ELISA Kit (E-EL-H2665c) were purchased from Elabscience, Wuhan, China.

Techniques:

Journal: eBioMedicine

Article Title: Reprogramming astrocytic NDRG2/NF-κB/C3 signaling restores the diabetes-associated cognitive dysfunction

doi: 10.1016/j.ebiom.2023.104653

Figure Lengend Snippet: NDRG2 reshapes the pathological structure of astrocytes by inhibiting NF-κB/C3 signaling. (a) GSEA of proteome indicated complement and coagulation cascade signaling were upregulated in diabetic mice (∗∗∗ p = 0.0002 between vehicle and STZ) and downregulated after exercise (∗∗ p = 0.0035 between STZ and STZ + Run). The cumulative enrichment scores were normalized (NES) ( n = 3/group). (b) Heatmaps showing the fold changes of significantly altered proteins in complement and coagulation cascades ( n = 3/group). (c – d) Representative immunofluorescent staining of intact C3 and its cleaved products in the hippocampus. Red: C3, green: GFAP, blue: DAPI. Scale bar, 50 μm. (∗∗∗ p = 0.0006 between vehicle and STZ, ∗∗∗ p = 0.0006 between STZ and STZ + Run) (e – f) Representative immunoblots of complement C3 in the hippocampus were increased in the STZ group compared to vehicle mice (∗∗∗∗ p < 0.0001 between vehicle and STZ), which was downregulated after exercise ( n = 6/group) (∗∗∗ p = 0.0008 between STZ and STZ + Run). (g – k) Representative immunoblots of astrocytic NDRG2 ( g, i ), complement C3 ( g, h ), NF-κB, and p–NF–κB ( j, k ) after NDRG2 loss of function ( n = 6/group) (∗ p = 0.0120 (h) , ∗∗∗∗ p < 0.0001 (i) , ∗∗ p = 0.0012 (k) between STZ + Run + acNDRG2 KO and STZ + Run + control). (l – n) Representative immunoblots of astrocytic NDRG2 ( l ), complement C3 ( m ), NF-κB, and p–NF–κB ( n ) in the hippocampus after NDRG2 gain of function ( n = 6/group) (∗∗ p = 0.0064 ( l ), ∗∗∗ p = 0.0001 ( m ), ∗∗ p = 0.0029 ( n ) between vehicle + AAV-Ctrl and STZ + AAV-Ctrl) (∗∗∗∗ p < 0.0001 ( l ), ∗ p = 0.0264 (m) , ∗∗ p = 0.0040 (n) between STZ + AAV-NDRG2 and STZ + AAV-Ctrl). Data are presented as mean ± SEM. GSEA analysis was performed in a . One-way ANOVA with Tukey's multiple comparisons test was performed in d, f, l–n . Two-tailed Student's t-test was performed in h, i, k .

Article Snippet: The Human C3 (Complement Component 3) ELISA Kit (E-EL-H6054) and Human INS (Insulin) ELISA Kit (E-EL-H2665c) were purchased from Elabscience, Wuhan, China.

Techniques: Coagulation, Staining, Western Blot, Control, Two Tailed Test

Journal: eBioMedicine

Article Title: Reprogramming astrocytic NDRG2/NF-κB/C3 signaling restores the diabetes-associated cognitive dysfunction

doi: 10.1016/j.ebiom.2023.104653

Figure Lengend Snippet: C3aR blockade rescues dendritic spine loss in diabetic mice, and C3 may be a biomarker to predict the progression of DACD in humans. (a) Schematic representing chronological order of STZ injection, C3aR antagonist, and behavioral testing. We used C3aR antagonists to clarify whether C3aR blockade could mimic the protective effect of NDRG2 overexpression on DACD. (b) Y-maze alternation triplet (%) was improved in the STZ + C3aRA group compared with STZ + PBS mice ( n = 7/group, n = 8 vehicle + PBS) (∗∗∗ p = 0.0006 between vehicle + PBS and STZ + PBS) (∗∗∗∗ p < 0.0001 between STZ + C3aRA and STZ + PBS). (c – d) Y-maze total distance ( c ) and total arm entries ( d ) ( n = 7/group, n = 8 vehicle + PBS). (e) Escape latency of MWM test was shorten at the STZ + C3aRA group compared with STZ + PBS mice ( n = 7/group, n = 8 vehicle + PBS) (∗ p = 0.0211 (day 1) , ∗ p = 0.0336 (day 2) , ∗∗ p = 0.0099 (day 3) , ∗∗ p = 0.0033 (day 4) between vehicle + PBS and STZ + PBS) ( ## p = 0.0021 (day 1) , # p = 0.0392 (day 3) between STZ + C3aRA and STZ + PBS). (f) Platform crossover of MWM test was upregulated at the STZ + C3aRA mice compared to STZ + PBS group ( n = 7/group, n = 8 vehicle + PBS) (∗∗ p = 0.0025 between vehicle + PBS and STZ + PBS) (∗∗ p = 0.0066 between STZ + C3aRA and STZ + PBS). (g) Representative images of 3D reconstruction of dendritic spines. Scale bar, 5 μm. (h – l) The densities of total spines ( h ), stubby spines ( i ), mushroom spines ( j ), long thin spines ( k ), and filopodia spines ( l ) ( n = 20/group) (∗∗∗∗ p < 0.0001 ( h ), ∗∗∗ p = 0.0008 ( i ), ∗ p = 0.0191 ( j ), ∗∗∗ p = 0.0006 ( k ) between vehicle + PBS and STZ + PBS) (∗∗∗∗ p < 0.0001 ( h ), ∗∗∗ p = 0.0005 ( i ), ∗ p = 0.0133 ( j ), ∗ p = 0.0153 ( l ) between STZ + C3aRA and STZ + PBS). (m) The DSST scores of diabetic patients ( n = 27) and non-diabetic peers ( n = 13) (∗∗∗∗ p < 0.0001 between Normal and Diabetes). (n) Fasting plasma glucose levels for diabetic patients ( n = 27) compared to non-diabetic patients ( n = 13) (∗∗∗∗ p < 0.0001 between Normal and Diabetes). (o) The serum levels of complement C3 in diabetic patients ( n = 27) and normal peers ( n = 13) (∗∗ p = 0.0072 between Normal and Diabetes). (p) The correlations between DSST score and increased C3 levels in diabetic patients ( n = 27) and normal peers ( n = 13). Correlations were found using linear regression, with r = −0.3315 and p = 0.0366. Values presented as mean ± SEM. Two-way ANOVA with Tukey's multiple comparisons test was performed in e . One-way ANOVA with Tukey's multiple comparisons test was performed in b, f–l . Two-tailed Student's t-test was performed in m–o .

Article Snippet: The Human C3 (Complement Component 3) ELISA Kit (E-EL-H6054) and Human INS (Insulin) ELISA Kit (E-EL-H2665c) were purchased from Elabscience, Wuhan, China.

Techniques: Biomarker Discovery, Injection, Over Expression, Clinical Proteomics, Two Tailed Test

Journal: eBioMedicine

Article Title: Reprogramming astrocytic NDRG2/NF-κB/C3 signaling restores the diabetes-associated cognitive dysfunction

doi: 10.1016/j.ebiom.2023.104653

Figure Lengend Snippet: Characteristics of the study population.

Article Snippet: The Human C3 (Complement Component 3) ELISA Kit (E-EL-H6054) and Human INS (Insulin) ELISA Kit (E-EL-H2665c) were purchased from Elabscience, Wuhan, China.

Techniques:

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: ELISA Kit for Complement C3 , Elabscience , E-EL-R0250.

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

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Cyclic-di-AMP Phosphodiesterase Elicits Protective Immune Responses Against Mycobacterium tuberculosis H37Ra Infection in Mice

doi: 10.3389/fcimb.2022.871135

Figure Lengend Snippet: Antibody and complement detection of CnpB-immunized mice after M. tuberculosis intranasal infection. After M. tuberculosis H37Ra intranasal challenge for 8 weeks, antibodies and complements were detected. (A) Sera of CnpB immunized (CnpB) mice and unimmunized (UN) mice were collected for antibodies detection using ELISA ( n =3). The collection time points was 8 weeks after infection. (B) The ratio of IgG2a/IgG1 in panel A was calculated ( n =3). (C, D) The levels of complement C3/C5 in sera ( n = 3) (C) and complement C3 in lung/spleen homogenate ( n = 4) (D) were assayed by ELISA. (E) The levels of sIgA in BALF were assayed by ELISA. The results are shown as mean ± SD. * P < 0.05, ** P < 0.01, **** P < 0.0001.

Article Snippet: Complement C3 and C5 were measured by the C3 ELISA kit (Alpha Diagnostic International, USA) and the C5 ELISA kit (Cloud-Clone Corp, USA) according to the manufacturer’s instructions.

Techniques: Infection, Enzyme-linked Immunosorbent Assay