Journal: bioRxiv

Article Title: Cell-derived ECM loaded electrospun Polycaprolactone/Chitosan nanofibrous scaffolds for periodontal regeneration

doi: 10.1101/2023.03.30.534964

Figure Lengend Snippet: Characterization of decellularized extracellular matrix (dECM) derived from human periodontal ligament stem cells (PDLSCs). A) Immunofluorescence staining images of fibronectin (FIB, green), collagen I (COL I, red), laminin (LAM, red), asporin (ASP, red), osteopontin (OPN, red) and osteocalcin (OC, red) before and after decellularization DAPI staining (blue) revealed the absence of nuclei after decellularization. Scale bar 100 μm. B) DNA content present in PDLSCs and dECM. C) Quantification of sulphated glycosaminoglycans (sGAGs) in PDLSCs and dECM. D) Content of collagen present in PDLSCs and dECM. Three different samples were used in each quantification assay (N=3) for both conditions; *** p < 0.001.

Article Snippet: The primary antibodies, including rabbit polyclonal collagen I (1:200, Abcam), rabbit polyclonal asporin (1:100, Thermo Fisher Scientific), mouse monoclonal osteopontin (1:100, Thermo Fisher Scientific), mouse monoclonal osteocalcin (1:50, R&D Systems), rabbit polyclonal periostin (1:100, Abcam) and rabbit polyclonal cementum protein 1 (1:100, Thermo Fisher Scientific) prepared in a solution of 0.3% Triton X-100, 1% BSA, 10% FBS (in PBS) were added into the samples and incubated overnight at 4°C.

Techniques: Derivative Assay, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Cell-derived ECM loaded electrospun Polycaprolactone/Chitosan nanofibrous scaffolds for periodontal regeneration

doi: 10.1101/2023.03.30.534964

Figure Lengend Snippet: Immunofluorescent staining images of collagen I (COL I, red), asporin (ASP, red), osteopontin (OPN, red), osteocalcin (OC, red), periostin (POSTN, red) and cementum protein 1 (CMP, red) expressed by PDLSCs cultured on PCL, PCL-CTS and PCL-CTS-ECM electrospun scaffolds for 21 days under osteogenic differentiation conditions. Nuclei were counterstained with DAPI (blue). Scale bar 100 μm.

Article Snippet: The primary antibodies, including rabbit polyclonal collagen I (1:200, Abcam), rabbit polyclonal asporin (1:100, Thermo Fisher Scientific), mouse monoclonal osteopontin (1:100, Thermo Fisher Scientific), mouse monoclonal osteocalcin (1:50, R&D Systems), rabbit polyclonal periostin (1:100, Abcam) and rabbit polyclonal cementum protein 1 (1:100, Thermo Fisher Scientific) prepared in a solution of 0.3% Triton X-100, 1% BSA, 10% FBS (in PBS) were added into the samples and incubated overnight at 4°C.

Techniques: Staining, Cell Culture

Journal: Cancer Cell International

Article Title: Silencing Uracil-DNA glycosylase inhibits colorectal cancer progression

doi: 10.1186/s12935-025-04089-y

Figure Lengend Snippet: UNG activated several signaling pathways in CRC cells. Western blot was performed to detect the levels of mTOR, p-mTOR, p70 S6K, p-P70 S6K, AKT, p-AKT, AMPK, p-AMPK, ERK, p-ERK, Bax, Bcl2, cleavages of caspase-9, and caspase-3 in UNG-knockdown CRC cells. Fold changes (Fc) are shown below the bars

Article Snippet: The membranes were incubated overnight at 4 °C with the following primary antibodies: UNG, P70 S6K, p-P70 S6K(S424) (1:1000, Bioworld), β-actin (1:1000, Abcam), cleaved caspase-3, AMPKα1/AMPKα2, p-AMPKα1(Thr183)/AMPKα2(Thr172) (1:1000, Beyotime), BAX, BCL2, AKT1/2/3, mTOR, and p-mTOR (Ser2448) (1:1000; BOSTER), cleaved caspase-9, p-AKT(Ser473), ERK1/2, and p-ERK1/2(Thr202/Tyr204)/(Thr185/Tyr187) (1:1000; ZEN BIO), and GAPDH (1:1000; Proteintech).

Techniques: Protein-Protein interactions, Western Blot, Knockdown

Journal: EBioMedicine

Article Title: Chlamydia trachomatis Prevents Apoptosis Via Activation of PDPK1-MYC and Enhanced Mitochondrial Binding of Hexokinase II

doi: 10.1016/j.ebiom.2017.08.005

Figure Lengend Snippet: CTL2 induces PDPK1 phosphorylation downstream of PI3K. a) Western blot showing an increase of PDPK1 phosphorylated at Ser-241 compared to total PDPK1 at 24 and 48 h p.i. in CTL2-infected (MOI 1) whole cell lysates. β-actin served as loading control. b) The band densities from a) were quantified and normalized to corresponding band densities of the β-actin loading control. Alterations in expression levels compared to non-infected controls are represented as mean fold change ± SEM, * p < 0.05, t -test, n = 3. c) CTL2 infection increases levels of PDPK1 phosphorylated at Ser-241, which is recruited as a rim-like structure at the inclusions in HeLa cells. PDPK1 phosphorylated at Ser-241, IncA and nuclei were labelled with Cy3-conjugated phosphorylated PDPK1 and IncA antibodies and DAPI. Scale bar: 30 μm. Asterisks are CTL2 inclusions. d) Monolayers of fallopian tube mesenchymal stem cells, infected with CTL2 for 48 h p.i. (MOI 0.5) were labelled with antibodies against Chlamydia trachomatis , PDPK1 phosphorylated at Ser-241 and DAPI. Results were similar to c). Scale bar: 30 μm. e) Human primary fallopian tube epithelial organoids, infected with CTL2 for 48 h p.i. were labelled with antibodies against Chlamydia trachomatis , PDPK1 phosphorylated at Ser-241 and E -Cadherin. Results were similar to c). Scale bar: 30 μm.

Article Snippet: The following antibodies were used: rabbit monoclonal antibodies against total MYC (D84C12, 5605), total HKII (C64G5, 2867) and total PDHK1 (C47H1, 3820), rabbit polyclonal antibodies against total PDPK1 (3062) and anti-P-PDPK1 (Ser-241, 3061) (all Cell Signaling); rabbit polyclonal anti-P-MYC (T58, ab28842), mouse monoclonal anti-P-MYC (S62, 33A12E10, ab78318, both from Abcam); mouse monoclonal anti-VDAC1 (B-6, sc-390996), Rho A (24C4, sc-418) and anti-MYC (9E10, sc-40 all from Santa Cruz Biotechnology Inc.); mouse monoclonal anti- Chlamydia trachomatis MAb species-specific KK-12 IgG2a (supplied by David Grayston, University of Washington, Seattle); rabbit polyclonal anti- Chlamydia genus-specific antibody (AG, 3-090, Milan Analytica), mouse monoclonal anti- Chlamydia trachomatis Hsp60 (ALX-804-072-R100, Enzo Life Sciences), rabbit anti-INCA antibody ( ); goat anti-chlamydia (1990-0404, AbD Serotec); rabbit monoclonal cleaved PARP (Asp214) and rabbit polyclonal anti-cleaved caspase 3 (Asp175, 9661, both from Cell Signaling).

Techniques: Phospho-proteomics, Western Blot, Infection, Control, Expressing