Journal: Bioactive Materials

Article Title: An intelligent nanoliposome alleviates disc degeneration and discogenic pain by inhibiting neurovascular ingrowth via a “Soil-conditioning, seed-modulating, and weeds-suppressing” strategy

doi: 10.1016/j.bioactmat.2025.11.048

Figure Lengend Snippet: NM-LP TK /RSV-MnCDs alleviates discogenic pain in IVDD. (A) Experimental timeline schematic. (B–C) Open-field behavioral analysis showing locomotor activity parameters (active time, central mean speed, total distance, and mean speed) across groups (n = 5). (D–E) PWL and PWT test showing alleviation of thermal and mechanical allodynia by NM-LP TK /RSV-MnCDs in LSI mice (n = 5). (F–G) IHC staining and quantification analyses of DRG sections showing expression of sensory neuropeptides CGRP and SP (n = 5). Scale bar, 100 μm. (H–I) IF co-labeling of PGP9.5 and CGRP/SP in DRG, and corresponding quantification analyses (n = 5). Scale bar, 100 μm. Data are presented as mean ± SD. For comparisons among multiple groups at a single time point, one-way ANOVA followed was employed to determine statistical significance. For datasets involving multiple groups evaluated over time, including PWL and PWT assessments, a two-way repeated measures ANOVA was utilized. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; ns = not significant.

Article Snippet: For immunofluorescence (IF) staining, antigen retrieval was conducted using citrate buffer (0.1 M, pH 6.0), followed by blocking in 10 % normal goat serum (Solarbio, Cat. No. SL038) at room temperature for 1 h. Sections were incubated overnight at 4 °C with primary antibodies targeting ACAN (Cat. No. DF7561, Affinity), COL2A1 (Cat. No. AF0135, Affinity), MMP3 (Cat. No. AF0217, Affinity), MMP13 (Cat. No. 18165-1-AP, Proteintech), CD31 (Cat. No. AF0077, Affinity), p-LATS1 (Cat. No. AF7170, Affinity), p-YAP (Cat. No. AF3328, Affinity), Tuj1 (Cat. No. 66375, Proteintech), CGRP (Cat. No. DF7386, Affinity), SP (Cat. No. DF7522, Affinity), and PGP9.5 (Cat. No. AF5490, Affinity) followed by incubation with appropriate fluorescent secondary antibodies for 2 h at room temperature and counterstaining with DAPI (Beyotime, Cat. No. C1006).

Techniques: Activity Assay, Immunohistochemistry, Expressing, Labeling

Journal: The Journal of Biological Chemistry

Article Title: Calcitonin gene-related peptide inhibits macrophage migration and differentiation via the GTPase Rap1

doi: 10.1016/j.jbc.2025.110949

Figure Lengend Snippet: BIBN alleviates CGRP-induced inhibition of macrophage migration. A, representative images of the scratch assay showing macrophage migration in the control, CGRP, BIBN, and BIBN + CGRP groups at 0 h and 24 h. The red dashed lines indicate the wound edges. This scale bar represents = 500 μm. Quantification of scratch (n = 3/group) on the right . B, representative images of the Transwell migration assay showing migrated macrophages in the control, CGRP, BIBN, and BIBN + CGRP groups. Cells that migrated through the membrane were stained with crystal violet . This scale bar represents = 100 μm. Quantification of Transwell assays (n = 7/group) on the right . C, representative images of phalloidin staining of BMMs cultured in the control, CGRP, BIBN, and BIBN + CGRP groups for 24 h, The scale bar represents = 20 μm; shape factor(n = 6/group)and length(n = 7/group). D, representative images showing F-actin ( red ) and Vinculin ( green ) staining of BMMs cultured in control, CGRP, BIBN, and BIBN + CGRP groups for 24 h, This scale bar represents = 10 μm. This scale bar represents in zoom = 1.5 μm; white arrows indicate filopodia (F-actin) and adhesion sites (Vinculin). E, quantification of filopodia and cell adhesion site is shown on the right . n = 6/group. F, expression of Rap1, PI3k, p-PI3k, Akt, p-Akt was examined by Western blot. β-actin was used as a loading control. Quantitative analyses of the relative band intensities of Rap1 to β-actin and p-AKTto AKT (n = 3). The p values were calculated by one-way analysis of variance. All data are presented as mean ± SD.

Article Snippet: To further investigate the effects of CGRP treatments on signaling pathways, 5 μm PI3K/AKT-IN-1 (Cat#HY-144806, MedChem-Express) was used to inhibit the PI3K-Akt signaling pathway.

Techniques: Inhibition, Migration, Wound Healing Assay, Control, Transwell Migration Assay, Membrane, Staining, Cell Culture, Expressing, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Calcitonin gene-related peptide inhibits macrophage migration and differentiation via the GTPase Rap1

doi: 10.1016/j.jbc.2025.110949

Figure Lengend Snippet: siRNA depletion of Rap1 reversed CGRP-induced phenotypes in macrophages. A, representative images of the scratch assay showing macrophage migration in the negative control (NC), negative control + CGRP, sirap1and sirap1+CGRP groups at 0 h and 24 h. The red dashed lines indicate the wound edges. This scale bar represents = 500 μm. Quantification of scratch (n = 3/group) on the right . B, representative images of the Transwell migration assay showing migrated macrophages in the negative control, negative control + CGRP, sirap1and sirap1+CGRP groups. Cells that migrated through the membrane were stained with crystal violet . This scale bar represents = 100 μm. Quantification of Transwell assays (n = 7/group) on the right . C, representative images of phalloidin staining of BMMs cultured in media containing negative control, negative control + CGRP, sirap1and sirap1+CGRP for 24 h, this scale bar represents = 20 μm; shape factor and length. n = 7. D, representative images showing F-actin ( red ) and Vinculin ( green ) staining of BMMs. This scale bar represent = 10 μm. This scale bar represents in zoom =1.5 μm. White arrows indicate filopodia (F-actin) and adhesion sites (Vinculin). E, quantification of filopodia and cell adhesion sites. n = 7/group. F, expression of E-cadherin, Vimentin, N-cadherin, PI3K, p-PI3K, p-AKT, AKT was examined by Western blot, GAPDH was used as a loading control. G, quantitative analyses of the relative band intensities of E-cadherin, N-cadherin, and Vimentin to GAPDH and p-PI3K to PI3K, p-AKT to AKT (n = 3). The p values were calculated by one-way analysis of variance ( A , B , C , and E ) or two-way analysis of variance ( G ). All data are presented as mean ± SD. CGRP, calcitonin gene-related peptide.

Article Snippet: To further investigate the effects of CGRP treatments on signaling pathways, 5 μm PI3K/AKT-IN-1 (Cat#HY-144806, MedChem-Express) was used to inhibit the PI3K-Akt signaling pathway.

Techniques: Wound Healing Assay, Migration, Negative Control, Transwell Migration Assay, Membrane, Staining, Cell Culture, Expressing, Western Blot, Control

Journal: The Journal of Biological Chemistry

Article Title: Calcitonin gene-related peptide inhibits macrophage migration and differentiation via the GTPase Rap1

doi: 10.1016/j.jbc.2025.110949

Figure Lengend Snippet: The PI3K/AKT inhibitor further suppressed macrophage migration. A, representative images of the scratch assay showing macrophage migration in the control, CGRP, PI3K/AKT-IN-1, PI3K/AKT-IN-1+CGRP groups at 0 h and 24 h. The red dashed lines indicate the wound edges. The scale bar represnts = 500 μm. Quantification of scratch (n = 3/group) on the right . B, representative images of the Transwell migration assay showing migrated macrophages in the control, CGRP, PI3K/AKT-IN-1, PI3K/AKT-IN-1+CGRP groups. Cells that migrated through the membrane were stained with crystal violet . Scale bar = 100 μm. Quantification of Transwell assays(n = 8/group) on the right . C, representative images of phalloidin staining of BMMs cultured in control, CGRP, PI3K/AKT-IN-1, PI3K/AKT-IN-1+CGRP groups for 24 h, This scale bar represent= 20 μm; shape factor, and length (n = 8). D , representative images showing F-actin ( red ) and Vinculin ( green ) staining of BMMs cultured in control, CGRP, PI3K/AKT-IN-1, PI3K/AKT-IN-1+CGRP groups for 24 h, This scale bar represents = 10 μm. This scale bar represents in zoom =1.5 μm. White arrow s indicate filopodia (F-actin) and adhesion sites (Vinculin). Quantification of filopodia and cell adhesion site is shown on the right . n = 8. The p values were calculated by one-way analysis of variance. All data are presented as mean ± SD. CGRP, calcitonin gene-related peptide.

Article Snippet: To further investigate the effects of CGRP treatments on signaling pathways, 5 μm PI3K/AKT-IN-1 (Cat#HY-144806, MedChem-Express) was used to inhibit the PI3K-Akt signaling pathway.

Techniques: Migration, Wound Healing Assay, Control, Transwell Migration Assay, Membrane, Staining, Cell Culture