Journal: Journal of Cellular and Molecular Medicine

Article Title: Integrating Bulk RNA and Single‐Cell RNA Sequencing Identifies and Validates Lactylation‐Related Signatures for Intervertebral Disc Degeneration

doi: 10.1111/jcmm.70262

Figure Lengend Snippet: Immune characteristics and interaction function of LRGs. (A) The bar plot shows the distribution of 20 distinct immune cells across various IVDD samples. (B) The box plot illustrates the expression profiles of 20 immune cells in the IVDD group and the control group. (C–H) Correlations of C IGFBP3, D CBX3, E THUMPD1, F CHST1, G DDIT4 and H RBM10 with immune cell infiltration.

Article Snippet: After incubation with 3% BSA for 25 min, the sections were then treated with primary antibody against CBX3 (11650‐2‐AP; Proteintech, Chian, 1:100) at 4°C for 12 h. The next day, the sections were exposed to secondary antibody (511203; ZENBIO, China, 1:300) for 60 min. Stained section images were observed using light microscopy (BX43; Olympus, Japan).

Techniques: Expressing, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Integrating Bulk RNA and Single‐Cell RNA Sequencing Identifies and Validates Lactylation‐Related Signatures for Intervertebral Disc Degeneration

doi: 10.1111/jcmm.70262

Figure Lengend Snippet: Validation of the expression of six hub LRGs. (A) Relative mRNA expression levels of six LRGs of NPCs treated with different dose IL‐1β (0, 10 30 50 ng/mL, 24 h). (B) UMAP visualisation displayed the expression of CBX3 in the MDD and SDD groups. (C, D) IHC staining and qualification analysis showing the expression of CBX3 in human disc degeneration tissues. Scale bar = 20 μm. (E, F) Representative images of HE staining, SF staining and histological scores in sham and AFP groups. Scale bar = 200 μm. (G, H) IHC staining and qualification analysis of CBX3 in the sham and AFP groups. Scale bar = 200 μm. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.000.

Article Snippet: After incubation with 3% BSA for 25 min, the sections were then treated with primary antibody against CBX3 (11650‐2‐AP; Proteintech, Chian, 1:100) at 4°C for 12 h. The next day, the sections were exposed to secondary antibody (511203; ZENBIO, China, 1:300) for 60 min. Stained section images were observed using light microscopy (BX43; Olympus, Japan).

Techniques: Biomarker Discovery, Expressing, Immunohistochemistry, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Integrating Bulk RNA and Single‐Cell RNA Sequencing Identifies and Validates Lactylation‐Related Signatures for Intervertebral Disc Degeneration

doi: 10.1111/jcmm.70262

Figure Lengend Snippet: Atosiban acetate as effective molecule alleviated NPC degeneration via repressing the glycolysis activity and global lactylation level. (A) Relative mRNA expression levels of CBX3 in NPCs treated with different molecules (10 μM, 24 h). (B) Relative mRNA expression levels of ACAN, COL2A1, ADAMTS5, MMP3, LDHA and PKM2 in NPCs after dilazep dihydrochloride, cobicistat, atosiban acetate and felypressin acetate. (C) Molecular docking of CBX3 and atosiban acetate. (D) Western blot analysis and quantification showing the protein expression of ACAN, COL2A1, ADAMTS5, MMP3, LDHA and PKM2 in NPCs in response to IL‐1β and atosiban acetate. (E) Relative mRNA expression levels of ACAN, COL2A1, ADAMTS5, MMP3, LDHA and PKM2 in NPCs in response to IL‐1β and atosiban acetate. (F) Western blot analysis showing the protein expression of Pan Kla in NPCs in response to IL‐1β and atosiban acetate. (G) IF staining showing the expression of Pan Kla in NPCs in response to IL‐1β and atosiban acetate. Scale bar = 20 μm. (H, I) Representative images of HE staining, SF staining and histological scores in different groups. Scale bar = 200 μm. (J, K) IHC staining and qualification analysis of ACAN and MMP3 in different groups. Scale bar = 200 μm. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001.

Article Snippet: After incubation with 3% BSA for 25 min, the sections were then treated with primary antibody against CBX3 (11650‐2‐AP; Proteintech, Chian, 1:100) at 4°C for 12 h. The next day, the sections were exposed to secondary antibody (511203; ZENBIO, China, 1:300) for 60 min. Stained section images were observed using light microscopy (BX43; Olympus, Japan).

Techniques: Activity Assay, Expressing, Western Blot, Staining, Immunohistochemistry