Journal: JBMR Plus

Article Title: Proteomic analysis and effects on osteogenic differentiation of exosomes from patients with ossification of the spinal ligament

doi: 10.1093/jbmrpl/ziaf021

Figure Lengend Snippet: (A) Western blots revealing Runx2 and Wnt3a expression at wk 2 and 3 following culture in osteogenic differentiation medium using exosome-penetrating (1.2 μm) and exosome-non-penetrating (0.03 μm) filters. (B) Expression levels of Runx2 and Wnt3a of OLF-derived and non-OLF-derived (control) cells co-cultured with 0.03- and 1.2-μm filters. ( n = 3 each). ** p <.01.

Article Snippet: The membranes were washed twice in PBS solution containing 0.05% Tween 20, then blocked by a mixture of 5% skimmed milk in PBS for 1 h at room temperature, and finally incubated overnight with one of the following antibodies at 4 °C: rabbit anti-Runx2 polyclonal Ab (dilution, 1:250, Proteintech) and rabbit anti-Wnt3a polyclonal Ab (dilution, 1:250, Proteintech) to assess the osteogenic differentiation, and rabbit anti-COLIVA1 polyclonal Ab (dilution, 1:250, GeneTex) rabbit anti-FMNL3 polyclonal Ab (dilution, 1:250, Proteintech), rabbit anti-mTOR polyclonal Ab (dilution, 1:250, Proteintech), or rabbit anti-PIP4K2B polyclonal Ab (dilution, 1:250, Proteintech) to confirm the differential expression of candidate proteins.

Techniques: Western Blot, Expressing, Derivative Assay, Control, Cell Culture

Journal: JBMR Plus

Article Title: Proteomic analysis and effects on osteogenic differentiation of exosomes from patients with ossification of the spinal ligament

doi: 10.1093/jbmrpl/ziaf021

Figure Lengend Snippet: (A) Western blots revealing Runx2 and Wnt3a expression at wk 2 and 3 following culture in osteogenic differentiation medium using exosome-penetrating (1.2 μm) and exosome-non-penetrating (0.03 μm) filters. (B) Expression levels of Runx2 and Wnt3a of OLF-derived and non-OLF-derived (control) cells co-cultured with 0.03- and 1.2-μm filters. ( n = 3 each). ** p <.01.

Article Snippet: The membranes were washed twice in PBS solution containing 0.05% Tween 20, then blocked by a mixture of 5% skimmed milk in PBS for 1 h at room temperature, and finally incubated overnight with one of the following antibodies at 4 °C: rabbit anti-Runx2 polyclonal Ab (dilution, 1:250, Proteintech) and rabbit anti-Wnt3a polyclonal Ab (dilution, 1:250, Proteintech) to assess the osteogenic differentiation, and rabbit anti-COLIVA1 polyclonal Ab (dilution, 1:250, GeneTex) rabbit anti-FMNL3 polyclonal Ab (dilution, 1:250, Proteintech), rabbit anti-mTOR polyclonal Ab (dilution, 1:250, Proteintech), or rabbit anti-PIP4K2B polyclonal Ab (dilution, 1:250, Proteintech) to confirm the differential expression of candidate proteins.

Techniques: Western Blot, Expressing, Derivative Assay, Control, Cell Culture

Journal: Heliyon

Article Title: Multi-omic analyses identified SFRP4 as a novel biomarker in abnormal uterine bleeding with ovulatory dysfunction.

doi: 10.1016/j.heliyon.2024.e37168

Figure Lengend Snippet: Fig. 7. Effects of SFRP4 knockdown in hEECs on the Wnt pathway. Expression of SFRP2, Wnt-3A, Wnt-5A, Wnt-7A and Fzd-5 was measured in hEECs transfected with si-NC or si-SFRP4 by western blotting 72 h later. (A) Representative western blots of SFRP2, Wnt-3A, Wnt-5A, Wnt-7A and Fzd-5 in the si-NC and si-SFRP4 groups (n = 4). For the uncropped versions of western blotting, please refer to Supplement Files. (B) Relative densitometric values of SFRP2, Wnt-3A, Wnt-5A, Wnt-7A and Fzd-5 in the si-NC and si-SFRP4 groups. Data are presented as mean ± SD. ns, no significance; *p < 0.05; **p < 0.01.

Article Snippet: Western blotting was conducted as previously done [17] The primary antibodies: rabbit antibody against SFRP4 at dilution 1:1000 (Invitrogen, USA), mouse antibody against GAPDH at dilution 1:10000 (Abcam, UK), rabbit antibody against SFRP2 at dilution 1:1000 (Abcam, UK), rabbit antibody against Wnt-3A at dilution 1:1000 (Cell Signaling, USA), rabbit antibody against Wnt-5A at dilution 1:500 (Abcam, UK), rabbit antibody against Wnt-7A at dilution 1:1000 (Abcam, UK), rabbit antibody against Fzd-5 at dilution 1:1000 (Abcam, UK).

Techniques: Knockdown, Expressing, Transfection, Western Blot