Journal: Cellular & Molecular Biology Letters

Article Title: The promoting roles of GLP1R and GIPR in stemness maintenance and multiple lineage-specific differentiation of PDLSCs

doi: 10.1186/s11658-026-00867-2

Figure Lengend Snippet: Cell growth of PDLSCs enhanced by GIPRA. a Western blot assay validate the expression of GLP1R and GIPR in PDLSCs treated with siRNA-mediated knockdown, ectopic overexpression, and agonists of GLP1R and GIPR. The interpretation of bubble plot is illustrated in Fig. a. b CCK-8 assay indicating the cell growth enhanced by GIPR agonist, but suppressed by GIPR knockdown ( n = 5 in each group). The interpretation of line colors in the line chart is the same in Fig. a. c Colony formation assay showing the crystal violet spots in different groups of PDLSCs (top). Histogram shows the statistical analysis of colony counts by Student’s t -test (bottom) ( n = 5 in each group). The interpretation of column colors is the same as in Fig. a. d Stacked column chart show the proportion of cells at different stages of cell cycle. * Significant increase of G2/M phase in GIPR KD and DKD groups as well as decrease of G2/M phase in GIPRA, 2GRA, and BGM0504 ( P < 0.05) compared with NC determined by Student’s t -test ( n = 5 in each group). e Venn diagram show the intersection of differentially expressed genes (|log 2 FC > 1|, P < 0.05) among the GIPR KD, GIPRA, and 2GRA groups compared with NC group. f The bubble plot displaying the top ten functions of cell proliferation (GO biological process) associated with the 1151 differentially expressed genes in e . g GSEA identifies the activated enrichment of the MAPK/ERK signaling pathway in response to GIPRA treatment compared with NC group. NC, normal PDLSCs as negative control; KD, knockdown; OE, overexpression; DKD, GLP1R and GIPR double knockdown; DOE, GLP1R and GIPR double overexpression; GLP1RA, GLP1R agonist; GIPRA, GIPR agonist; 2GRA, GLP1R/GIPR agonist-1; BGM0504, BGM0504 injection of GLP-1 and GIP dual receptor agonist (developed by BrightGene)

Article Snippet: The in vitro treatment conditions for the small-molecule compound were as follows: GLP1RA semaglutide acetate (10 nM, 24 h), GIPRA (Pro3) GIP (0.5 nM, 24 h), GLP-1R/GIPR agonist-1 (10 nM, 24 h) also termed as 2GRA in this study, BGM0504 (2 nM, 24 h, BrightGene Pharmaceutical Co., Ltd., China) [ ], CREB inhibitor 666-15 (1 μM, 2 h, HY-101120, MCE), Akt inhibitor MK-2206 (5 μM, 24 h, HY-108232, MCE), ERK1/2 inhibitor SCH772984 (0.5 μM, 24 h, HY-50846, MCE), β-catenin inhibitor IN-3 (5 μM, 24 h, HY-147007, MCE), STAT3-IN-14 (5 μM, 2 h, HY-N10472, MCE).

Techniques: Western Blot, Expressing, Knockdown, Over Expression, CCK-8 Assay, Colony Assay, Negative Control, Injection

Journal: Cellular & Molecular Biology Letters

Article Title: The promoting roles of GLP1R and GIPR in stemness maintenance and multiple lineage-specific differentiation of PDLSCs

doi: 10.1186/s11658-026-00867-2

Figure Lengend Snippet: Translation shut off by upregulated IFIT proteins. a Heatmap showing the differentially expressed genes (|log 2 FC|> 1, P < 0.05) compared between GLP1R knockdown, GLP1RA, 2GRA, and NC among the multiple lineage differentiation system of PDLSCs. b–e Dumbbell plots illustrating the changes in gene expression of IFIT1 , IFIT2 , IFIT3 , IRF9 , IFI44 , IFI6 , IFIH1 , OAS1 , OASL , RSAD2 , and BST2 between GLP1R knockdown, GLP1RA, 2GRA, and NC in osteogenesis ( b ), adipogenesis ( c ), chondrogenesis ( d ), and neurogenesis ( e ) from RNA sequencing data. f Western blot assay showing the expression of classic markers of osteogenesis COL1A1, BGLAP, and RUNX2 in PDLSCs for osteogenic differentiation system treated with GLP1R knockdown, GLP1RA, and 2GRA. g Western blot assay showing the expression of classic markers of adipogenesis CEBPA, FABP4, and PPARG in PDLSCs for adipogenic differentiation system treated with GLP1R knockdown, GLP1RA, and 2GRA. h Western blot assay show the expression of classic markers of chondrogenesis ACAN, COL2A1, and SOX9 in PDLSCs for chondrogenic differentiation system treated with GLP1R knockdown, GLP1RA, and 2GRA. i Western blot assay show the expression of classic markers of neurogenesis MAP2, NES, and TUBB3 in PDLSCs for adipogenic differentiation system treated with GLP1R knockdown, GLP1RA, and 2GRA. j Western blot assay showing the interaction of IFIT1 with IFIT2, IFIT3, eIF3C, and eIF3E within the immunoprecipitation of IFIT1 from PDLSCs under undifferentiated, osteogenic, GLP1R knockdown, GLP1RA, and 2GRA treated conditions. k Western blot assay show the interaction of eIF3C with IFIT1, IFIT2, IFIT3, eIF3E, and RPS3 within the immunoprecipitation of eIF3C from PDLSCs under undifferentiated, osteogenic, GLP1R knockdown, GLP1RA, and 2GRA treated conditions. Red boxes highlight the robust protein interaction in undifferentiated PDLSCs and osteogenic PDLSCs with GLP1R knockdown. KD, knockdown; NC, PDLSCs for regular differentiation accordingly; Un, undifferentiated PDLSCs; OS, osteogenesis; 2GRA, GLP1R/GIPR agonist-1

Article Snippet: The in vitro treatment conditions for the small-molecule compound were as follows: GLP1RA semaglutide acetate (10 nM, 24 h), GIPRA (Pro3) GIP (0.5 nM, 24 h), GLP-1R/GIPR agonist-1 (10 nM, 24 h) also termed as 2GRA in this study, BGM0504 (2 nM, 24 h, BrightGene Pharmaceutical Co., Ltd., China) [ ], CREB inhibitor 666-15 (1 μM, 2 h, HY-101120, MCE), Akt inhibitor MK-2206 (5 μM, 24 h, HY-108232, MCE), ERK1/2 inhibitor SCH772984 (0.5 μM, 24 h, HY-50846, MCE), β-catenin inhibitor IN-3 (5 μM, 24 h, HY-147007, MCE), STAT3-IN-14 (5 μM, 2 h, HY-N10472, MCE).

Techniques: Knockdown, Gene Expression, RNA Sequencing, Western Blot, Expressing, Immunoprecipitation

Journal: Cellular & Molecular Biology Letters

Article Title: The promoting roles of GLP1R and GIPR in stemness maintenance and multiple lineage-specific differentiation of PDLSCs

doi: 10.1186/s11658-026-00867-2

Figure Lengend Snippet: The protective effect of PDLSCs by GLP1RA and GIPRA from periodontitis in vivo. a Representative bidimensional views of the maxillary molars of PD mice model treated with GLP1RA and GIPRA by micro-CT. Scale bar, 0.5 mm. Measurement of the distance from CEJ to ABC around second molar. Histogram shows the differential distance among different groups. Red columns indicate a significant increase compared with NC (gray column), whereas yellow columns indicate a significant decrease compared with red ( P < 0.05) determined by Student’s t -test ( n = 5 in each group). b Representative images of TRAP staining of the maxillary molars of those groups from ( a ). Scale bar, 20 μm. Measurement of the osteoclast number around second molar. Histogram show the differential distance among different groups. c Representative bidimensional views of the maxillary molars of PD mice model treated with GLP1RA, GIPRA, and PDLSC transplantation by micro-CT ( n = 5 in each group). Scale bar, 0.5 mm. Measurement of the distance from CEJ to ABC around second molar. Histogram shows the differential distance among different groups. Green columns indicate a significant decrease compared with NC (gray columns). d Representative images of TRAP staining of the maxillary molars of those groups from c . Scale bar, 20 μm. Measurement of the osteoclast number around second molar. Histogram shows the differential distance among different groups. e – i Immunofluorescence shows mCherry fluorescence derived from exogenous PDLSCs ( n = 5 in each group). The fluorescence indicated the expression of genes such as CD146 (e), RUNX2 (f), CD29 ( g ), UCP1 ( h ), and IFIT1 ( i ), thereby enabling the evaluation of the proliferation and multilineage differentiation of the transplanted PDLSCs under different treatment conditions. Scale bar, 20 μm. WT, wild type; KO, GLP1R −/− /GIPR −/− ; PD, periodontitis; PDLSCs, PDLSCs transplantation; GLP1RA, GLP1R agonist; GIPRA, GIPR agonist; 2GRA, GLP1R/GIPR agonist-1; CEJ, cemento-enamel junction; ABC, alveolar bone crest

Article Snippet: The in vitro treatment conditions for the small-molecule compound were as follows: GLP1RA semaglutide acetate (10 nM, 24 h), GIPRA (Pro3) GIP (0.5 nM, 24 h), GLP-1R/GIPR agonist-1 (10 nM, 24 h) also termed as 2GRA in this study, BGM0504 (2 nM, 24 h, BrightGene Pharmaceutical Co., Ltd., China) [ ], CREB inhibitor 666-15 (1 μM, 2 h, HY-101120, MCE), Akt inhibitor MK-2206 (5 μM, 24 h, HY-108232, MCE), ERK1/2 inhibitor SCH772984 (0.5 μM, 24 h, HY-50846, MCE), β-catenin inhibitor IN-3 (5 μM, 24 h, HY-147007, MCE), STAT3-IN-14 (5 μM, 2 h, HY-N10472, MCE).

Techniques: In Vivo, Micro-CT, Staining, Transplantation Assay, Immunofluorescence, Fluorescence, Derivative Assay, Expressing