Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

Article Title: Association of the α 2 δ 1 Subunit with Ca v 3.2 Enhances Membrane Expression and Regulates Mechanically Induced ATP Release in MLO-Y4 Osteocytes

doi: 10.1002/jbmr.437

Figure Lengend Snippet: RT-PCR primers used for detection of VSCC subunits and osteocyte markers

Article Snippet: The rabbit anti-β 3 antibody was purchased from Alomone Research Laboratories (Jerusalem, Israel).

Techniques: Sequencing

Journal: International immunopharmacology

Article Title: Inhibiting the SARM1-NAD + axis reduces oxidative stress-induced damage to retinal and nerve cells.

doi: 10.1016/j.intimp.2024.112193

Figure Lengend Snippet: Fig. 6. DSRM ameliorated oxidative stress-induced neurite injury in N2a cells. (A-C) GOx inhibited N2a cell proliferation and neurite growth in the RA-induced N2a cell differentiation model (magnification 400 × ); (D-E) Immunocytochemistry demonstrated that 10 μM DSRM pre-treatment for 2 h enhanced β3-tubulin expression induced by 7 mU/mL GOx for 12 h in N2a cells (scale bar: 50 μm). N ≥3, **p < 0.01, ****p < 0.0001.

Article Snippet: After culturing and treating the cells in 24-well plates and fixing and permeabilizing them with methanol at -20 ◦C for 15 min, the cells were then washed 3 times with PBS, and then incubated in containment buffer (5 % donkey serum for containment) for 20 min, and then incubated with anti-GSDMD primary antibody (1:500, Abclonal, Wuhan, China, Cat#A20197), anti-β3-tubulin primary antibody (1:1000, CST, USA, Cat#5568S) was incubated at 4 ◦C overnight.

Techniques: Cell Differentiation, Immunocytochemistry, Expressing

Journal: Prion

Article Title: Role of Prion protein-EGFR multimolecular complex during neuronal differentiation of human dental pulp-derived stem cells

doi: 10.1080/19336896.2018.1463797

Figure Lengend Snippet: Differentiation of hDPSCs. (A) Morphology of hDPSCs from dental pulp untreated and treated with EGF/bFGF for 14 days. Scale bars, 50 μm. (B) Immunofluorescence analysis of neuronal marker β3-tubulin mAb expression. Scale bars, 50 μm.

Article Snippet: After washing, cells were incubated with rabbit anti-PrP EP1802Y mAb for 1 h at 4 °C, followed by CY5-conjugated anti-rabbit IgG H&L (Abcam, Cambridge, USA) for additional 30 min. Alternatively we performed a double staining with rabbit anti-PrP EP1802Y mAb and mouse anti-CD44 mAb or mouse anti-β3-tubulin mAb (Cell Signaling Technology, Danvers, MA, USA).

Techniques: Immunofluorescence, Marker, Expressing

Journal: Prion

Article Title: Role of Prion protein-EGFR multimolecular complex during neuronal differentiation of human dental pulp-derived stem cells

doi: 10.1080/19336896.2018.1463797

Figure Lengend Snippet: PrPC expression in hDSPCs. (A) Flow cytometry analysis of PrPC expression at 21 and 28 days from dental pulp separation and after additional 7 and 14 days with EGF/bFGF. Histograms represent log fluorescence vs cell number, gated on cell population of a side scatter/forward scatter (SS/FS) histogram. Cell number is indicated on the y-axis and fluorescence intensity is represented on the x-axis. Each panel was compared with the corresponding IgG negative isotype control. A representative experiment among 3 is shown. (B) Double staining flow cytometry analysis of PrP/CD44 in control hDPSCs (28 days from pulp separation) and PrP/ β3-tubulin after stimulation with EGF/bFGF for additional 14 days. Histograms represent log fluorescence PE vs log fluorescence CY5, gated on cell population of a side scatter/forward scatter (SS/FS) histogram. CD44-PE fluorescence is indicated on the y-axis and PrP-CY5 fluorescence intensity is represented on the x-axis. Each panel was compared with the corresponding IgG negative isotype control. A representative experiment among 3 is shown. (C) Western blot analysis of PrPC expression at 21 and 28 days from dental pulp separation and after additional 7 and 14 days with EGF/bFGF, using anti-PrP SAF32. Loading control was evaluated using anti-β-actin. Densitometric analysis of bands from the representative western blot is reported in panel on the right as Mean ± SD. (D) Immunofluorescence analysis of hDPSC untreated o treated with EGF/bFGF, using anti-PrP SAF32.

Article Snippet: After washing, cells were incubated with rabbit anti-PrP EP1802Y mAb for 1 h at 4 °C, followed by CY5-conjugated anti-rabbit IgG H&L (Abcam, Cambridge, USA) for additional 30 min. Alternatively we performed a double staining with rabbit anti-PrP EP1802Y mAb and mouse anti-CD44 mAb or mouse anti-β3-tubulin mAb (Cell Signaling Technology, Danvers, MA, USA).

Techniques: Expressing, Flow Cytometry, Fluorescence, Control, Double Staining, Western Blot, Immunofluorescence

Journal: Prion

Article Title: Role of Prion protein-EGFR multimolecular complex during neuronal differentiation of human dental pulp-derived stem cells

doi: 10.1080/19336896.2018.1463797

Figure Lengend Snippet: Role of PrPC during neuronal differentiation of hDPSCs induced by EGF/bFGF. (A) Flow cytometry analysis of PrPC expression in hDPSCs, untreated or treated with siRNA PrP for 72 hours. Histograms represent log fluorescence vs cell number, gated on cell population of a side scatter/forward scatter (SS/FS) histogram. Cell number is indicated on the y-axis and fluorescence intensity is represented on the x-axis. Each panel was compared with the corresponding IgG negative isotype control. A representative experiment among 3 is shown. (B-C) Western blot analysis of β3-tubulin, and NFH expression in hDPSCs, untreated or treated with 20 ng/ml EGF and 40 ng/ml bFGF for 14 days, in the presence or in the absence of pre-treatment with siRNA PrP or scrambled siRNA for 72 hours. Densitometric analysis is shown in the right panel. Results represent the Mean±SD from 3 independent experiments. *p <0.01 siRNAPrP treated cells vs EGF/bFGF treated cells.

Article Snippet: After washing, cells were incubated with rabbit anti-PrP EP1802Y mAb for 1 h at 4 °C, followed by CY5-conjugated anti-rabbit IgG H&L (Abcam, Cambridge, USA) for additional 30 min. Alternatively we performed a double staining with rabbit anti-PrP EP1802Y mAb and mouse anti-CD44 mAb or mouse anti-β3-tubulin mAb (Cell Signaling Technology, Danvers, MA, USA).

Techniques: Flow Cytometry, Expressing, Fluorescence, Control, Western Blot