Journal: International Journal of Molecular Sciences

Article Title: Proanthocyanidins Ameliorate LPS-Inhibited Osteogenesis of PDLSCs by Restoring Lysine Lactylation

doi: 10.3390/ijms25052947

Figure Lengend Snippet: Osteogenesis and lactylation levels are decreased in inflamed periodontal tissue and LPS-stimulated PDLSCs. ( A ) H&E staining in rat periodontal tissues and immunohistochemical Pan Kla and RUNX2 expression. ( B ) Quantitative analysis of positive areas. ( C ) Western blotting showing protein levels of COL-1, ALP, RUNX2, and BMP2 in LPS-treated PDLSCs. ( D ) qRT-PCR showing mRNA levels of osteogenic genes COL-1 , ALP, RUNX2 , and BMP2 . ( E ) ALP staining at 7 days and ARS staining at 21 days of osteogenic induction in LPS-treated PDLSCs. ( F ) Lactate concentration in the culture medium of PDLSCs was measured at the indicated times, as shown in the quantitative analysis. ( G ) Western blotting showing the protein level of Pan Kla in LPS-treated PDLSCs. ( H ) Quantitative analysis of Pan Kla protein levels. ( I ) Immunofluorescence staining images showing the expression levels of RUNX2 (green) and Pan Kla (red) in PDLSCs after LPS treatment. Cell nuclei were stained with DAPI (blue). PDLSCs: periodontal ligament stem cells; Kla: lysine lactylation; LPS: lipopolysaccharide; NC: healthy mice without periodontitis; H&E: hematoxylin and eosin staining (Data are expressed as mean ± SD, N = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, NS indicates no statistical difference).

Article Snippet: Primary antibodies are as follows: rabbit ant-IL-1β (1:1000, Cell Signaling Technology, Danvers, MA, USA); rabbit anti-coll I antibody (1:1000, ABclonal, Wuhan, China); rabbit anti-ALP antibody (1:1000, ABclonal, Wuhan, China); rabbit anti-RUNX2 antibody (1:1000, ABclonal, Wuhan, China); rabbit anti-BMP2 antibody (1:1000, ABclonal, Wuhan, China); and rabbit anti-Pan Kla antibody (1:1000, PTM Bio, Hangzhou, China).

Techniques: Staining, Immunohistochemical staining, Expressing, Western Blot, Quantitative RT-PCR, Concentration Assay, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Proanthocyanidins Ameliorate LPS-Inhibited Osteogenesis of PDLSCs by Restoring Lysine Lactylation

doi: 10.3390/ijms25052947

Figure Lengend Snippet: Restoration of lactylation by TSA recovers the osteogenesis of PDLSCs in an inflammatory state. ( A ) Western blotting showing the protein levels of COL-1, ALP, RUNX2, and BMP2 in PDLSCs after treatment with the conditions as shown. ( B ) Quantification of protein levels of COL-1, ALP, RUNX2, and BMP2. ( C ) qRT-PCR showing mRNA levels of osteogenic genes COL-1 , ALP, RUNX2, and BMP2 . ( D ) ALP staining at 7 days and ARS staining at 21 days of osteogenic induction in PDLSCs. ( E ) Quantitative analysis of ALP staining and ARS staining. ( F ) Immunofluorescence staining images showing the expression levels of RUNX2 (green) and Pan Kla (red) in PDLSCs treated with the conditions as shown. Cell nuclei were stained with DAPI (blue). ( G ) Pearson linear regression analysis of RUNX2 and Pan Ka fluorescence intensity. PDLSCs: periodontal ligament stem cells; Kla: lysine lactylation; LPS: lipopolysaccharide; TSA: Trichostatin A (Data are expressed as mean ± SD, N = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, NS indicates no statistical difference).

Article Snippet: Primary antibodies are as follows: rabbit ant-IL-1β (1:1000, Cell Signaling Technology, Danvers, MA, USA); rabbit anti-coll I antibody (1:1000, ABclonal, Wuhan, China); rabbit anti-ALP antibody (1:1000, ABclonal, Wuhan, China); rabbit anti-RUNX2 antibody (1:1000, ABclonal, Wuhan, China); rabbit anti-BMP2 antibody (1:1000, ABclonal, Wuhan, China); and rabbit anti-Pan Kla antibody (1:1000, PTM Bio, Hangzhou, China).

Techniques: Western Blot, Quantitative RT-PCR, Staining, Immunofluorescence, Expressing, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Proanthocyanidins Ameliorate LPS-Inhibited Osteogenesis of PDLSCs by Restoring Lysine Lactylation

doi: 10.3390/ijms25052947

Figure Lengend Snippet: Proanthocyanidins promote osteogenesis and elevate the lactylation of PDLSCs. ( A ) Western blotting showing protein levels of COL-1, ALP, RUNX2, and BMP2 in PA-treated PDLSCs. ( B ) Quantification of protein levels of COL-1, ALP, RUNX2, and BMP2. ( C ) qRT-PCR showing mRNA levels of osteogenic genes COL-1 , ALP, RUNX2 , and BMP2 . ( D ) ALP staining at 7 days and ARS staining at 21 days of osteogenic induction in PA-treated PDLSCs. ( E ) Lactate concentration in the culture medium of PDLSCs was measured at the indicated times as shown in the quantitative analysis. ( F ) Western blotting showing the protein level of Pan Kla in PA-treated PDLSCs. ( G ) Quantitative analysis of Pan Kla protein levels. ( H ) Immunofluorescence staining images showing the expression levels of RUNX2 (green) and Pan Kla (red) in PDLSCs after treatment with the conditions as shown. Cell nuclei were stained with DAPI (blue). PDLSCs: periodontal ligament stem cells; Kla: lysine lactylation; PA: proanthocyanidins (Data are expressed as mean ± SD, N = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, NS indicates no statistical difference).

Article Snippet: Primary antibodies are as follows: rabbit ant-IL-1β (1:1000, Cell Signaling Technology, Danvers, MA, USA); rabbit anti-coll I antibody (1:1000, ABclonal, Wuhan, China); rabbit anti-ALP antibody (1:1000, ABclonal, Wuhan, China); rabbit anti-RUNX2 antibody (1:1000, ABclonal, Wuhan, China); rabbit anti-BMP2 antibody (1:1000, ABclonal, Wuhan, China); and rabbit anti-Pan Kla antibody (1:1000, PTM Bio, Hangzhou, China).

Techniques: Western Blot, Quantitative RT-PCR, Staining, Concentration Assay, Immunofluorescence, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Proanthocyanidins Ameliorate LPS-Inhibited Osteogenesis of PDLSCs by Restoring Lysine Lactylation

doi: 10.3390/ijms25052947

Figure Lengend Snippet: Proanthocyanidins recover PDLSC osteogenesis by restoring the lactylation of PDLSCs in an inflammatory state. ( A ) Western blotting showing the protein levels of COL-1, ALP, RUNX2, and BMP2 in PDLSCs after treatment with the conditions as shown. ( B ) Quantification of protein levels of COL-1, ALP, RUNX2, and BMP2. ( C ) qRT-PCR showing mRNA levels of osteogenic genes COL-1 , ALP, RUNX2, and BMP2 . ( D ) ALP staining at 7 days and ARS staining at 21 days of osteogenic induction in PDLSCs. ( E ) Quantitative analysis of ALP staining and ARS staining. ( F ) Immunofluorescence staining images showing the expression levels of RUNX2 (green) and Pan Kla (red) in PDLSCs treated with the conditions as shown. Cell nuclei were stained with DAPI (blue). ( G ) Pearson linear regression analysis of RUNX2 and Pan Ka fluorescence intensity. PDLSCs: periodontal ligament stem cells; Kla: lysine lactylation; PA: proanthocyanidins; Theo-24: theophylline (Data are expressed as mean ± SD, N = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, NS indicates no statistical difference).

Article Snippet: Primary antibodies are as follows: rabbit ant-IL-1β (1:1000, Cell Signaling Technology, Danvers, MA, USA); rabbit anti-coll I antibody (1:1000, ABclonal, Wuhan, China); rabbit anti-ALP antibody (1:1000, ABclonal, Wuhan, China); rabbit anti-RUNX2 antibody (1:1000, ABclonal, Wuhan, China); rabbit anti-BMP2 antibody (1:1000, ABclonal, Wuhan, China); and rabbit anti-Pan Kla antibody (1:1000, PTM Bio, Hangzhou, China).

Techniques: Western Blot, Quantitative RT-PCR, Staining, Immunofluorescence, Expressing, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Proanthocyanidins Ameliorate LPS-Inhibited Osteogenesis of PDLSCs by Restoring Lysine Lactylation

doi: 10.3390/ijms25052947

Figure Lengend Snippet: Primer sequences.

Article Snippet: Primary antibodies are as follows: rabbit ant-IL-1β (1:1000, Cell Signaling Technology, Danvers, MA, USA); rabbit anti-coll I antibody (1:1000, ABclonal, Wuhan, China); rabbit anti-ALP antibody (1:1000, ABclonal, Wuhan, China); rabbit anti-RUNX2 antibody (1:1000, ABclonal, Wuhan, China); rabbit anti-BMP2 antibody (1:1000, ABclonal, Wuhan, China); and rabbit anti-Pan Kla antibody (1:1000, PTM Bio, Hangzhou, China).

Techniques: