Journal: Oncotarget

Article Title: Genome-wide shRNA screen revealed integrated mitogenic signaling between dopamine receptor D2 (DRD2) and epidermal growth factor receptor (EGFR) in glioblastoma

doi:

Figure Lengend Snippet: (A) DRD2 antagonists decreased pERK accumulation in the U87MG glioblastoma cell line. D: DMSO. S: spiperone (5 μM), H: haloperidol (5 μM), R: risperidone (10 μM), L: L-741,626 (10 μM). Cells were treated with the various antagonists for 1 hr before lysis/immuno-blotting. (B) Decreased pERK activation by DRD2 shRNA can be rescued by expression of an RNAi resistant form of DRD2 (DRD2RR). U87MG lines harboring a doxycycline inducible shDRD2 construct and pCMV6-DRD2RR or pCMV6 were treated with vehicle or doxycycline (1 μg/ml) for 48 hrs before lysis. Bottom panel: quantitative ratio of pERK to ERK. (C) DRD2 antagonists decreased pERK accumulation in the patient-derived CMK3 GSC line in a dose-dependent manner. D: DMSO. S: spiperone. H: haloperidol. CMK3 was treated with the various antagonists for 1 hr before lysis. (D) Treatment with a DRD2 agonist, quinpirole, enhanced pERK accumulation (left upper). CMK3 was treated with quinpirole for 1 hr prior to lysis/immuno-blotting. Lower panel: quantitative ratio of pERK to ERK. Quinpirole promoted cellular growth in the CMK3 GSC line detected by CellTiter-Blue (right). Cells were treated with 10 μM quinpirole for 5 days. (E) qPCR analysis revealed increased GNAI2 mRNA expression in glioblastoma specimens relative to the surrounding normal brain. 5 matched normal-glioblastoma specimens are shown. N: normal brain; T: glioblastoma tumor specimens. GNAI2 mRNA level was normalized to 18S rRNA mRNA. Similar results were obtained when normalized to actin or GAPDH. (F) GNAI2 siRNA transfection compromised proliferation of U87MG cells (top) and reduced pERK accumulation (bottom). #1 and #2 denotes two independent siRNAs against GNAI2. Cells were seeded at ~50% confluency, transfected with siRNAs for 48 hrs, and re-plated in serial dilution for clonogenic survival assessment. (G) The pro-proliferative effect of quinpirole in the patient-derived 1123 GSC line was inhibited by GNAI2 silencing. Cells were transfected with siGNAI2s for 48 hrs prior to treatment with 10 μM quinpirole for 5 days. Viability was determined using CellTiter-Blue (Promega). (H) GNAI2 silencing inhibited quinpirole-induced pERK accumulation and reversed quinpirole-suppression of Rap1-GTP accumulation in U87MG cells. Cells were transfected with siRNAs for 48 hrs prior to the treatment with 10 μM quinpirole for 1 hr.

Article Snippet: Antibodies used included: DRD2 (1:1000, Millipore); pERK (1:2000), ERK (1:2000), pAKT473 (1:2000), pS6 (1:2000), and pSTAT3 (1:1000) from Cell Signaling Technology; GNAI2 (1:1000, Proteintech); Tubulin (1:5000, Sigma); Ku86 (1:1000, Santa Cruz Biotechnology).

Techniques: Lysis, Activation Assay, shRNA, Expressing, Construct, Derivative Assay, Transfection, Serial Dilution

Journal: Physiological Reports

Article Title: Acute tuft cell ablation in mice induces malabsorption and alterations in secretory and immune cell lineages in the small intestine

doi: 10.14814/phy2.70264

Figure Lengend Snippet: Primary antibody list.

Article Snippet: GIP , Rabbit poly , Unconjugated , Novus , NBP3‐04865 , 1:100.

Techniques: Conjugation Assay

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: Reduced expression of GLP1R and GIPR in PDLSCs under multiple stress conditions. a Western blot assay show the expression of GLP1R and GIPR and PKA activity in PDLSCs respectively treated with cigarette, E-cig, hypoxia, LPS, alcohol, HG as well as MOF stimulation. Bubble plot indicates the quantitative analysis of the grayscale values of each band normalized to the β-actin (All western blot assays in this study were performed three duplications). Green bubbles indicate a significant decrease, whereas red bubbles indicate a significant increase compared to NC (gray bubble) ( P < 0.05) determined by Student’s t -test. Gray bubbles in other groups indicate no statistical significance compared with NC. b–d FRET-based biosensors detect the intracellular levels of cAMP ( b ), Ca 2+ ( c ), and PKA activity ( d ). CFP and YFP ratiometric images are shown as a pseudocolored green (CFP) and magenta (YFP) overlay (top). Quantifications of CFP/YFP emission ratio in the panels of sensor in different groups ( n = 20 in each group) (bottom). A ratio value more than 1 represents increased cAMP level, decreased Ca 2+ level, or inactivated PKA. e Western blot assay showing the expression of GLP1R and GIPR in PDLSCs derived from five healthy donors and five patients with periodontitis. The interpretation of bubble plot is illustrated in Fig. 1a. f , g Immunofluorescence show green fluorescence expression of the classical marker STRO-1, which is used to locate PDLSCs, and simultaneously demonstrate the co-localized red fluorescence expression of GLP1R ( f ) and GIPR ( g ) in NC (top) and PD (bottom) in vivo. H&E staining (right panel) is first performed to guide the identification of periodontal ligament tissue at second molar of NC and PD in serial sections, followed by immunofluorescence detection. Scale bar, 20 μm. Histogram show the differential positive expression rate of GLP1R or GIPR in STRO-1 positive cells compared between NC and PD. Green column indicate a significant decrease compared with NC (gray column) ( P < 0.05) determined by Student’s t -test (five fields for each group). NC, normal PDLSCs as negative control; Cig, cigarette; E-cig, E-cigarette; LPS, lipopolysaccharide; HG, high glucose; MOF, mild orthodontic force; FRET, Förster resonance energy transfer; HC, healthy control; PD, periodontitis

Article Snippet: The information and the concentration of antibodies used in this study were listed as follows: Phospho-(Ser/Thr) PKA substrate antibody (1: 500, cat. no. 9621, CST, USA), GLP1R (1: 2,000, cat. no. 26196-1-AP, Proteintech, China), GIPR (1: 2,000, cat. no. 28322-1-AP, Proteintech), STRO-1 (1: 250, cat. no. 39-8401, Thermo Fisher Scientific), CD146 (1: 2,000, cat. no. 65181-1-Ig, Proteintech), Vimentin (1: 2,000, cat. no. 10366-1-AP, Proteintech), CREB1 (1: 2,000, cat. no. 67927-1-Ig, Proteintech), p-CREB1 (Ser133) (1: 2,000, cat. no. 28792-1-AP, Proteintech), ERK1/2 (1: 2,000, cat. no. 66192-1-Ig, Proteintech), p-ERK1/2 (Thr202/Tyr204) (1: 2,000, cat. no. 28733-1-AP, Proteintech), β-catenin (1: 2,000, cat. no. 51067-2-AP, Proteintech), p-β-catenin (Ser33) (1: 2,000, cat. no. 80067-1-RR, Proteintech), STAT3 (1: 2,000, cat. no. 10253-2-AP, Proteintech), p-STAT3 (Ser727) (1: 2,000, cat. no. 80199-2-RR, Proteintech), Ki-67 (1: 2,000, cat. no. 84192-4-RR, Proteintech), PCNA (1: 2,000, cat. no. 10205-2-AP, Proteintech), CCND3 (1: 2,000, cat. no. 26755-1-AP, Proteintech), β-actin (1: 2,000, cat. no. 66009-1- Ig, Proteintech), COL1A1 (1: 2,000, cat. no. 67288-1-Ig, Proteintech), OCN (1: 2,000, cat. no. 20277-1-AP, Proteintech), RUNX2 (1: 2,000, cat. no. 20700-1-AP, Proteintech), C/EBPα (1: 2,000, cat. no. 29388-1-AP, Proteintech), FABP4 (1: 2,000, cat. no. 12802-1-AP, Proteintech), PPARγ (1: 2,000, cat. no. 16643-1-AP, Proteintech), ACAN (1: 2,000, cat. no. 68350-1-Ig, Proteintech), COL2A1 (1: 2,000, cat. no. 28459-1-AP, Proteintech), SOX9 (1: 2,000, cat. no. 67439-1-Ig, Proteintech), MAP2 (1: 2,000, cat. no. 17490-1-AP, Proteintech), Nestin (1: 2,000, cat. no. 19483-1-AP, Proteintech), TUBB3 (1: 2,000, cat. no. 66375-1-Ig, Proteintech), IFIT1 (1: 2,000, cat. no. ab305301, Abcam), IFIT2 (1: 2,000, cat. no. 12604-1-AP, Proteintech), IFIT3 (1: 2,000, cat. no. 15201-1-AP, Proteintech), eIF3C (1: 2,000, cat. no. 12733-1-AP, Proteintech), eIF3E (1: 2,000, cat. no. 10899-1-AP, Proteintech), RPS3 (1: 2,000, cat. no. 66046-1-Ig, Proteintech), SPP1 (1: 2,000, cat. no. 22952-1-AP, Proteintech), FGF18 (1: 2,000, cat. no. 11495-1-AP, Proteintech), MMP14 (1: 2,000, cat. no. 14552-1-AP, Proteintech), BMP2 (1: 2,000, cat. no. 66383-1-Ig, Proteintech), horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (1: 5,000, cat. no. SA00001-2, Proteintech), and HRP-conjugated goat anti-mouse IgG (1: 5,000, cat. no. SA00001-1, Proteintech).

Techniques: Expressing, Western Blot, Activity Assay, Derivative Assay, Immunofluorescence, Fluorescence, Marker, In Vivo, Staining, Negative Control, Förster Resonance Energy Transfer, Control

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 information and the concentration of antibodies used in this study were listed as follows: Phospho-(Ser/Thr) PKA substrate antibody (1: 500, cat. no. 9621, CST, USA), GLP1R (1: 2,000, cat. no. 26196-1-AP, Proteintech, China), GIPR (1: 2,000, cat. no. 28322-1-AP, Proteintech), STRO-1 (1: 250, cat. no. 39-8401, Thermo Fisher Scientific), CD146 (1: 2,000, cat. no. 65181-1-Ig, Proteintech), Vimentin (1: 2,000, cat. no. 10366-1-AP, Proteintech), CREB1 (1: 2,000, cat. no. 67927-1-Ig, Proteintech), p-CREB1 (Ser133) (1: 2,000, cat. no. 28792-1-AP, Proteintech), ERK1/2 (1: 2,000, cat. no. 66192-1-Ig, Proteintech), p-ERK1/2 (Thr202/Tyr204) (1: 2,000, cat. no. 28733-1-AP, Proteintech), β-catenin (1: 2,000, cat. no. 51067-2-AP, Proteintech), p-β-catenin (Ser33) (1: 2,000, cat. no. 80067-1-RR, Proteintech), STAT3 (1: 2,000, cat. no. 10253-2-AP, Proteintech), p-STAT3 (Ser727) (1: 2,000, cat. no. 80199-2-RR, Proteintech), Ki-67 (1: 2,000, cat. no. 84192-4-RR, Proteintech), PCNA (1: 2,000, cat. no. 10205-2-AP, Proteintech), CCND3 (1: 2,000, cat. no. 26755-1-AP, Proteintech), β-actin (1: 2,000, cat. no. 66009-1- Ig, Proteintech), COL1A1 (1: 2,000, cat. no. 67288-1-Ig, Proteintech), OCN (1: 2,000, cat. no. 20277-1-AP, Proteintech), RUNX2 (1: 2,000, cat. no. 20700-1-AP, Proteintech), C/EBPα (1: 2,000, cat. no. 29388-1-AP, Proteintech), FABP4 (1: 2,000, cat. no. 12802-1-AP, Proteintech), PPARγ (1: 2,000, cat. no. 16643-1-AP, Proteintech), ACAN (1: 2,000, cat. no. 68350-1-Ig, Proteintech), COL2A1 (1: 2,000, cat. no. 28459-1-AP, Proteintech), SOX9 (1: 2,000, cat. no. 67439-1-Ig, Proteintech), MAP2 (1: 2,000, cat. no. 17490-1-AP, Proteintech), Nestin (1: 2,000, cat. no. 19483-1-AP, Proteintech), TUBB3 (1: 2,000, cat. no. 66375-1-Ig, Proteintech), IFIT1 (1: 2,000, cat. no. ab305301, Abcam), IFIT2 (1: 2,000, cat. no. 12604-1-AP, Proteintech), IFIT3 (1: 2,000, cat. no. 15201-1-AP, Proteintech), eIF3C (1: 2,000, cat. no. 12733-1-AP, Proteintech), eIF3E (1: 2,000, cat. no. 10899-1-AP, Proteintech), RPS3 (1: 2,000, cat. no. 66046-1-Ig, Proteintech), SPP1 (1: 2,000, cat. no. 22952-1-AP, Proteintech), FGF18 (1: 2,000, cat. no. 11495-1-AP, Proteintech), MMP14 (1: 2,000, cat. no. 14552-1-AP, Proteintech), BMP2 (1: 2,000, cat. no. 66383-1-Ig, Proteintech), horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (1: 5,000, cat. no. SA00001-2, Proteintech), and HRP-conjugated goat anti-mouse IgG (1: 5,000, cat. no. SA00001-1, Proteintech).

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: Cell differentiation of PDLSCs enhanced by GLP1RA. a Representative images of Alizarin Red S staining of mineralized nodules in PDLSCs by osteogenic induction and siRNA-mediated knockdown and ectopic overexpression of GLP1R or GIPR as well as agonists of GLP1R and GIPR. Calcium deposits are visualized as red-stained areas, indicating extracellular matrix mineralization. Quantification was performed by dye extraction and absorbance measurement at 562 nm. Scar bar, 100 μm. b Representative images of Oil Red O staining in PDLSCs cultured by adipogenic induction and siRNA-mediated knockdown and ectopic overexpression of GLP1R or GIPR as well as agonists of GLP1R and GIPR. Red staining indicate intracellular lipid droplet accumulation. Quantification was performed by dye extraction and absorbance measurement at 490 nm. Scale bar, 100 μm. c Representative images of Alcian Blue staining in PDLSCs cultured by chondrogenic induction and siRNA-mediated knockdown and ectopic overexpression of GLP1R or GIPR as well as agonists of GLP1R and GIPR. Blue coloration indicates the deposition of sulfated glycosaminoglycans, a hallmark of chondrogenic differentiation. Quantification was performed by dye extraction and absorbance measurement at 620 nm. Scale bar, 100 μm. d Double immunofluorescence staining show the co-localization of NeuN (Red) and GFAP (Green) representing neurons and astrocytes in PDLSCs by neurogenic induction and siRNA-mediated knockdown and ectopic overexpression of GLP1R or GIPR as well as agonists of GLP1R and GIPR. Scar bar, 50 μm. Histogram shows the differential percentages of NeuN or GFAP positive cells among these groups (five fields for per group) determined by one-way ANOVA. e Western blot assay show the expression of STRO-1, CD146, and vimentin as mesenchymal biomarkers of PDLSCs by osteogenic induction with siRNA-mediated knockdown and ectopic overexpression of GLP1R or GIPR as well as agonists of GLP1R and GIPR. In statistical analysis, the meanings of the red, green, and gray colors are illustrated in Fig. a ( n = 5 in each group). Interpretation of abbreviations can be found in the legend of Fig.

Article Snippet: The information and the concentration of antibodies used in this study were listed as follows: Phospho-(Ser/Thr) PKA substrate antibody (1: 500, cat. no. 9621, CST, USA), GLP1R (1: 2,000, cat. no. 26196-1-AP, Proteintech, China), GIPR (1: 2,000, cat. no. 28322-1-AP, Proteintech), STRO-1 (1: 250, cat. no. 39-8401, Thermo Fisher Scientific), CD146 (1: 2,000, cat. no. 65181-1-Ig, Proteintech), Vimentin (1: 2,000, cat. no. 10366-1-AP, Proteintech), CREB1 (1: 2,000, cat. no. 67927-1-Ig, Proteintech), p-CREB1 (Ser133) (1: 2,000, cat. no. 28792-1-AP, Proteintech), ERK1/2 (1: 2,000, cat. no. 66192-1-Ig, Proteintech), p-ERK1/2 (Thr202/Tyr204) (1: 2,000, cat. no. 28733-1-AP, Proteintech), β-catenin (1: 2,000, cat. no. 51067-2-AP, Proteintech), p-β-catenin (Ser33) (1: 2,000, cat. no. 80067-1-RR, Proteintech), STAT3 (1: 2,000, cat. no. 10253-2-AP, Proteintech), p-STAT3 (Ser727) (1: 2,000, cat. no. 80199-2-RR, Proteintech), Ki-67 (1: 2,000, cat. no. 84192-4-RR, Proteintech), PCNA (1: 2,000, cat. no. 10205-2-AP, Proteintech), CCND3 (1: 2,000, cat. no. 26755-1-AP, Proteintech), β-actin (1: 2,000, cat. no. 66009-1- Ig, Proteintech), COL1A1 (1: 2,000, cat. no. 67288-1-Ig, Proteintech), OCN (1: 2,000, cat. no. 20277-1-AP, Proteintech), RUNX2 (1: 2,000, cat. no. 20700-1-AP, Proteintech), C/EBPα (1: 2,000, cat. no. 29388-1-AP, Proteintech), FABP4 (1: 2,000, cat. no. 12802-1-AP, Proteintech), PPARγ (1: 2,000, cat. no. 16643-1-AP, Proteintech), ACAN (1: 2,000, cat. no. 68350-1-Ig, Proteintech), COL2A1 (1: 2,000, cat. no. 28459-1-AP, Proteintech), SOX9 (1: 2,000, cat. no. 67439-1-Ig, Proteintech), MAP2 (1: 2,000, cat. no. 17490-1-AP, Proteintech), Nestin (1: 2,000, cat. no. 19483-1-AP, Proteintech), TUBB3 (1: 2,000, cat. no. 66375-1-Ig, Proteintech), IFIT1 (1: 2,000, cat. no. ab305301, Abcam), IFIT2 (1: 2,000, cat. no. 12604-1-AP, Proteintech), IFIT3 (1: 2,000, cat. no. 15201-1-AP, Proteintech), eIF3C (1: 2,000, cat. no. 12733-1-AP, Proteintech), eIF3E (1: 2,000, cat. no. 10899-1-AP, Proteintech), RPS3 (1: 2,000, cat. no. 66046-1-Ig, Proteintech), SPP1 (1: 2,000, cat. no. 22952-1-AP, Proteintech), FGF18 (1: 2,000, cat. no. 11495-1-AP, Proteintech), MMP14 (1: 2,000, cat. no. 14552-1-AP, Proteintech), BMP2 (1: 2,000, cat. no. 66383-1-Ig, Proteintech), horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (1: 5,000, cat. no. SA00001-2, Proteintech), and HRP-conjugated goat anti-mouse IgG (1: 5,000, cat. no. SA00001-1, Proteintech).

Techniques: Cell Differentiation, Staining, Knockdown, Over Expression, Extraction, Cell Culture, Double Immunofluorescence Staining, Western Blot, Expressing

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 information and the concentration of antibodies used in this study were listed as follows: Phospho-(Ser/Thr) PKA substrate antibody (1: 500, cat. no. 9621, CST, USA), GLP1R (1: 2,000, cat. no. 26196-1-AP, Proteintech, China), GIPR (1: 2,000, cat. no. 28322-1-AP, Proteintech), STRO-1 (1: 250, cat. no. 39-8401, Thermo Fisher Scientific), CD146 (1: 2,000, cat. no. 65181-1-Ig, Proteintech), Vimentin (1: 2,000, cat. no. 10366-1-AP, Proteintech), CREB1 (1: 2,000, cat. no. 67927-1-Ig, Proteintech), p-CREB1 (Ser133) (1: 2,000, cat. no. 28792-1-AP, Proteintech), ERK1/2 (1: 2,000, cat. no. 66192-1-Ig, Proteintech), p-ERK1/2 (Thr202/Tyr204) (1: 2,000, cat. no. 28733-1-AP, Proteintech), β-catenin (1: 2,000, cat. no. 51067-2-AP, Proteintech), p-β-catenin (Ser33) (1: 2,000, cat. no. 80067-1-RR, Proteintech), STAT3 (1: 2,000, cat. no. 10253-2-AP, Proteintech), p-STAT3 (Ser727) (1: 2,000, cat. no. 80199-2-RR, Proteintech), Ki-67 (1: 2,000, cat. no. 84192-4-RR, Proteintech), PCNA (1: 2,000, cat. no. 10205-2-AP, Proteintech), CCND3 (1: 2,000, cat. no. 26755-1-AP, Proteintech), β-actin (1: 2,000, cat. no. 66009-1- Ig, Proteintech), COL1A1 (1: 2,000, cat. no. 67288-1-Ig, Proteintech), OCN (1: 2,000, cat. no. 20277-1-AP, Proteintech), RUNX2 (1: 2,000, cat. no. 20700-1-AP, Proteintech), C/EBPα (1: 2,000, cat. no. 29388-1-AP, Proteintech), FABP4 (1: 2,000, cat. no. 12802-1-AP, Proteintech), PPARγ (1: 2,000, cat. no. 16643-1-AP, Proteintech), ACAN (1: 2,000, cat. no. 68350-1-Ig, Proteintech), COL2A1 (1: 2,000, cat. no. 28459-1-AP, Proteintech), SOX9 (1: 2,000, cat. no. 67439-1-Ig, Proteintech), MAP2 (1: 2,000, cat. no. 17490-1-AP, Proteintech), Nestin (1: 2,000, cat. no. 19483-1-AP, Proteintech), TUBB3 (1: 2,000, cat. no. 66375-1-Ig, Proteintech), IFIT1 (1: 2,000, cat. no. ab305301, Abcam), IFIT2 (1: 2,000, cat. no. 12604-1-AP, Proteintech), IFIT3 (1: 2,000, cat. no. 15201-1-AP, Proteintech), eIF3C (1: 2,000, cat. no. 12733-1-AP, Proteintech), eIF3E (1: 2,000, cat. no. 10899-1-AP, Proteintech), RPS3 (1: 2,000, cat. no. 66046-1-Ig, Proteintech), SPP1 (1: 2,000, cat. no. 22952-1-AP, Proteintech), FGF18 (1: 2,000, cat. no. 11495-1-AP, Proteintech), MMP14 (1: 2,000, cat. no. 14552-1-AP, Proteintech), BMP2 (1: 2,000, cat. no. 66383-1-Ig, Proteintech), horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (1: 5,000, cat. no. SA00001-2, Proteintech), and HRP-conjugated goat anti-mouse IgG (1: 5,000, cat. no. SA00001-1, Proteintech).

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 information and the concentration of antibodies used in this study were listed as follows: Phospho-(Ser/Thr) PKA substrate antibody (1: 500, cat. no. 9621, CST, USA), GLP1R (1: 2,000, cat. no. 26196-1-AP, Proteintech, China), GIPR (1: 2,000, cat. no. 28322-1-AP, Proteintech), STRO-1 (1: 250, cat. no. 39-8401, Thermo Fisher Scientific), CD146 (1: 2,000, cat. no. 65181-1-Ig, Proteintech), Vimentin (1: 2,000, cat. no. 10366-1-AP, Proteintech), CREB1 (1: 2,000, cat. no. 67927-1-Ig, Proteintech), p-CREB1 (Ser133) (1: 2,000, cat. no. 28792-1-AP, Proteintech), ERK1/2 (1: 2,000, cat. no. 66192-1-Ig, Proteintech), p-ERK1/2 (Thr202/Tyr204) (1: 2,000, cat. no. 28733-1-AP, Proteintech), β-catenin (1: 2,000, cat. no. 51067-2-AP, Proteintech), p-β-catenin (Ser33) (1: 2,000, cat. no. 80067-1-RR, Proteintech), STAT3 (1: 2,000, cat. no. 10253-2-AP, Proteintech), p-STAT3 (Ser727) (1: 2,000, cat. no. 80199-2-RR, Proteintech), Ki-67 (1: 2,000, cat. no. 84192-4-RR, Proteintech), PCNA (1: 2,000, cat. no. 10205-2-AP, Proteintech), CCND3 (1: 2,000, cat. no. 26755-1-AP, Proteintech), β-actin (1: 2,000, cat. no. 66009-1- Ig, Proteintech), COL1A1 (1: 2,000, cat. no. 67288-1-Ig, Proteintech), OCN (1: 2,000, cat. no. 20277-1-AP, Proteintech), RUNX2 (1: 2,000, cat. no. 20700-1-AP, Proteintech), C/EBPα (1: 2,000, cat. no. 29388-1-AP, Proteintech), FABP4 (1: 2,000, cat. no. 12802-1-AP, Proteintech), PPARγ (1: 2,000, cat. no. 16643-1-AP, Proteintech), ACAN (1: 2,000, cat. no. 68350-1-Ig, Proteintech), COL2A1 (1: 2,000, cat. no. 28459-1-AP, Proteintech), SOX9 (1: 2,000, cat. no. 67439-1-Ig, Proteintech), MAP2 (1: 2,000, cat. no. 17490-1-AP, Proteintech), Nestin (1: 2,000, cat. no. 19483-1-AP, Proteintech), TUBB3 (1: 2,000, cat. no. 66375-1-Ig, Proteintech), IFIT1 (1: 2,000, cat. no. ab305301, Abcam), IFIT2 (1: 2,000, cat. no. 12604-1-AP, Proteintech), IFIT3 (1: 2,000, cat. no. 15201-1-AP, Proteintech), eIF3C (1: 2,000, cat. no. 12733-1-AP, Proteintech), eIF3E (1: 2,000, cat. no. 10899-1-AP, Proteintech), RPS3 (1: 2,000, cat. no. 66046-1-Ig, Proteintech), SPP1 (1: 2,000, cat. no. 22952-1-AP, Proteintech), FGF18 (1: 2,000, cat. no. 11495-1-AP, Proteintech), MMP14 (1: 2,000, cat. no. 14552-1-AP, Proteintech), BMP2 (1: 2,000, cat. no. 66383-1-Ig, Proteintech), horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (1: 5,000, cat. no. SA00001-2, Proteintech), and HRP-conjugated goat anti-mouse IgG (1: 5,000, cat. no. SA00001-1, Proteintech).

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

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: Clinical impact of GLP1R and GIPR agonists on periodontitis severity. a , b Pie charts illustrating the distribution of periodontitis stage ( a ) and grade ( b ) in patients treated with GLP1R/GIPR agonists ( n = 74) (left) and those not treated ( n = 76) (right). ( c , d ) Box plots show the relationship between the duration of GLP1R/GIPR agonist administration and the severity of periodontitis, represented by stage ( c ) and grade ( d ). Data suggest a potential association between prolonged agonist use and reduced periodontitis severity. The classification of periodontitis stage and grade was assessed by the clinical attachment loss, probing pocket depth, and radiographic bone loss on the basis of the 2017 AAP/EFP classification

Article Snippet: The information and the concentration of antibodies used in this study were listed as follows: Phospho-(Ser/Thr) PKA substrate antibody (1: 500, cat. no. 9621, CST, USA), GLP1R (1: 2,000, cat. no. 26196-1-AP, Proteintech, China), GIPR (1: 2,000, cat. no. 28322-1-AP, Proteintech), STRO-1 (1: 250, cat. no. 39-8401, Thermo Fisher Scientific), CD146 (1: 2,000, cat. no. 65181-1-Ig, Proteintech), Vimentin (1: 2,000, cat. no. 10366-1-AP, Proteintech), CREB1 (1: 2,000, cat. no. 67927-1-Ig, Proteintech), p-CREB1 (Ser133) (1: 2,000, cat. no. 28792-1-AP, Proteintech), ERK1/2 (1: 2,000, cat. no. 66192-1-Ig, Proteintech), p-ERK1/2 (Thr202/Tyr204) (1: 2,000, cat. no. 28733-1-AP, Proteintech), β-catenin (1: 2,000, cat. no. 51067-2-AP, Proteintech), p-β-catenin (Ser33) (1: 2,000, cat. no. 80067-1-RR, Proteintech), STAT3 (1: 2,000, cat. no. 10253-2-AP, Proteintech), p-STAT3 (Ser727) (1: 2,000, cat. no. 80199-2-RR, Proteintech), Ki-67 (1: 2,000, cat. no. 84192-4-RR, Proteintech), PCNA (1: 2,000, cat. no. 10205-2-AP, Proteintech), CCND3 (1: 2,000, cat. no. 26755-1-AP, Proteintech), β-actin (1: 2,000, cat. no. 66009-1- Ig, Proteintech), COL1A1 (1: 2,000, cat. no. 67288-1-Ig, Proteintech), OCN (1: 2,000, cat. no. 20277-1-AP, Proteintech), RUNX2 (1: 2,000, cat. no. 20700-1-AP, Proteintech), C/EBPα (1: 2,000, cat. no. 29388-1-AP, Proteintech), FABP4 (1: 2,000, cat. no. 12802-1-AP, Proteintech), PPARγ (1: 2,000, cat. no. 16643-1-AP, Proteintech), ACAN (1: 2,000, cat. no. 68350-1-Ig, Proteintech), COL2A1 (1: 2,000, cat. no. 28459-1-AP, Proteintech), SOX9 (1: 2,000, cat. no. 67439-1-Ig, Proteintech), MAP2 (1: 2,000, cat. no. 17490-1-AP, Proteintech), Nestin (1: 2,000, cat. no. 19483-1-AP, Proteintech), TUBB3 (1: 2,000, cat. no. 66375-1-Ig, Proteintech), IFIT1 (1: 2,000, cat. no. ab305301, Abcam), IFIT2 (1: 2,000, cat. no. 12604-1-AP, Proteintech), IFIT3 (1: 2,000, cat. no. 15201-1-AP, Proteintech), eIF3C (1: 2,000, cat. no. 12733-1-AP, Proteintech), eIF3E (1: 2,000, cat. no. 10899-1-AP, Proteintech), RPS3 (1: 2,000, cat. no. 66046-1-Ig, Proteintech), SPP1 (1: 2,000, cat. no. 22952-1-AP, Proteintech), FGF18 (1: 2,000, cat. no. 11495-1-AP, Proteintech), MMP14 (1: 2,000, cat. no. 14552-1-AP, Proteintech), BMP2 (1: 2,000, cat. no. 66383-1-Ig, Proteintech), horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (1: 5,000, cat. no. SA00001-2, Proteintech), and HRP-conjugated goat anti-mouse IgG (1: 5,000, cat. no. SA00001-1, Proteintech).

Techniques:

Journal: The Journal of general virology

Article Title: Equine herpesvirus 1 infection orchestrates the expression of chemokines in equine respiratory epithelial cells.

doi: 10.1099/jgv.0.001317

Figure Lengend Snippet: Fig. 1. Expression patterns of CXCL9 and CXCL10 in mock-, abortigenic (N752) EHV1, or neurovirulent (D752) EHV1-inoculated ERECs. The left panel shows representative confocal images of the immunofluorescence staining, with (a) CXCL9 or (b) CXCL10 stained in green (FITC), viral proteins stained in red (TR), and nuclei counterstained in blue (Hoechst) in mock- or EHV1-inoculated ERECs, at 24 h p.i. Bars, 75 µm. The right panel shows the intensity of CXCL9 or CXCL10 signal in the ERECs at the level randomly chosen for viral plaques. Data are represented as means+sd, and ns indicates no statistically significant differences. *Statistically significant differences at P<0.05. Experiments were performed with ERECs isolated from three individual horses.

Article Snippet: CXCL9 and CXCL10 were visualized by incubating with a rabbit polyclonal anti-CXCL9 antibody (IgG; GeneTex) (1:50 in DPBS) or a rabbit polyclonal anti-CXCL10 antibody (IgG; Kingfisher Biotech) (1:50 in DPBS) followed by an FITCconjugated goat anti-rabbit IgG antibody (1:100 in DPBS) (Molecular Probes).

Techniques: Expressing, Immunofluorescence, Staining, Isolation

Journal: Frontiers in Microbiology

Article Title: Single-Molecule RNA Sequencing Reveals IFNγ-Induced Differential Expression of Immune Escape Genes in Merkel Cell Polyomavirus–Positive MCC Cell Lines

doi: 10.3389/fmicb.2021.785662

Figure Lengend Snippet: Overview of all differentially expressed genes categorized according to their activity in cancer biology.

Article Snippet: For antibody staining, 1 × 10 6 cells of each MCC cell line were cultured in a six-well plate in the presence or absence of IFNγ (3,000 U/ml) for 72 h. Cells were washed with fluorescence-activated cell sorting (FACS) buffer (dulbecco’s phosphate-buffered saline with 1% FBS and 0.2% sodium azide) and stained for the following markers: STAT1α/β, Indoleamine-2,3-dioxygenase (IDO), C-X-C motif chemokine 10 (CXCL10), PD-L1, bone marrow stromal antigen 2 (BST2), HLA class II histocompatibility antigen gamma chain (CD74), and HLA A, B, and C (HLA-ABC) with the following monoclonal antibodies: anti-human STAT1 (phycoerythine; PE, clone REA272, Miltenyi Biotec), anti-IDO1 (PE, clone eyedio, Invitrogen), anti-human CXCL10 (PE, clone REA334, Miltenyi Biotec), anti-CD274 (Pe-Cy7, clone MIH1, BD Biosciences), anti-human BST2 (PE, clone REA202, Miltenyi Biotec), anti-CD74 (PE, clone 5-329, Miltenyi Biotec), and anti–HLA-ABC (fluorescein-5-isothiocyanat, BD Biosciences).

Techniques: Activity Assay, Ubiquitin Proteomics

Journal: Frontiers in Microbiology

Article Title: Single-Molecule RNA Sequencing Reveals IFNγ-Induced Differential Expression of Immune Escape Genes in Merkel Cell Polyomavirus–Positive MCC Cell Lines

doi: 10.3389/fmicb.2021.785662

Figure Lengend Snippet: Validation of differential gene expression in MCC cell lines on protein level and mRNA level. (A) TPM values were acquired from our sequencing output. The data represent mean values ± SEM from three independent experiments (two for MKL-1 control). (B) After 72 h of treatment with or without IFNγ (3,000 U/ml), the Merkel cell carcinoma cell lines MKL-1, MKL-2, and WaGa were stained for signal transducer and activator of transcription 1-alpha/beta (STAT1), bone marrow stromal antigen 2 (BST2), C-X-C motif chemokine 10 (CXCL10), and HLA class II histocompatibility antigen gamma chain (CD74) and analyzed by flow cytometry. The average MFI of three independent experiments ± standard error is indicated. p -values were determined using the paired student’s t -test. * p < 0.05; ** p < 0.01; ns, not significant.

Article Snippet: For antibody staining, 1 × 10 6 cells of each MCC cell line were cultured in a six-well plate in the presence or absence of IFNγ (3,000 U/ml) for 72 h. Cells were washed with fluorescence-activated cell sorting (FACS) buffer (dulbecco’s phosphate-buffered saline with 1% FBS and 0.2% sodium azide) and stained for the following markers: STAT1α/β, Indoleamine-2,3-dioxygenase (IDO), C-X-C motif chemokine 10 (CXCL10), PD-L1, bone marrow stromal antigen 2 (BST2), HLA class II histocompatibility antigen gamma chain (CD74), and HLA A, B, and C (HLA-ABC) with the following monoclonal antibodies: anti-human STAT1 (phycoerythine; PE, clone REA272, Miltenyi Biotec), anti-IDO1 (PE, clone eyedio, Invitrogen), anti-human CXCL10 (PE, clone REA334, Miltenyi Biotec), anti-CD274 (Pe-Cy7, clone MIH1, BD Biosciences), anti-human BST2 (PE, clone REA202, Miltenyi Biotec), anti-CD74 (PE, clone 5-329, Miltenyi Biotec), and anti–HLA-ABC (fluorescein-5-isothiocyanat, BD Biosciences).

Techniques: Biomarker Discovery, Gene Expression, Sequencing, Control, Staining, Flow Cytometry