Journal: Journal of Orthopaedic Translation

Article Title: Comparative proteomic analysis identifies differentially expressed proteins and reveals potential mechanisms of traumatic heterotopic ossification progression

doi: 10.1016/j.jot.2022.04.003

Figure Lengend Snippet: qPCR and immunoblot assay compared the differential expression of the UBA1, EIF3E, RPL27, RPS28, RPL17 in tissue samples of rat models and clinical specimens. The relative gene expression of UBA1, EIF3E, RPL27, RPS28, RPL17 was evaluated in (a) clinical specimens and (b) tissue samples of AT, PAT and control rat models using qRT-PCR (c, d) Western blots for UBA1, EIF3E, RPL27, RPS28, RPL17 on whole-tissue lysate from clinical specimens and control, AT, PAT rat models. ​β-Actin served as a loading control. The relative expression level of UBA1, EIF3E, RPL27, RPS28, RPL17 in (e) clinical specimens and (f) tissue samples of control, AT, PAT rat models quantified using ImageJ and normalized to b-actin is shown. All tests in triplicate, ∗ represent significant change in the expression level compared to control group, ∗P ​< ​0.05, ∗∗P ​< ​0.01.

Article Snippet: Then, the slides were incubated with antibodies against UBA1(1:100, ab180125, Abcam Inc, Cambridge, UK), EIF3E (1:50, ab134958, Abcam Inc, Cambridge, UK), RPL27 (1:50, 14980-1-AP, Proteintech, Chicago, USA), RPL17 (1:300, 67223-1-Ig, Proteintech, Chicago, USA), RPS28(1:50, 14796-1-AP, Proteintech, Chicago, USA) at 4 °C overnight.

Techniques: Western Blot, Quantitative Proteomics, Gene Expression, Control, Quantitative RT-PCR, Expressing

Journal: Journal of Orthopaedic Translation

Article Title: Comparative proteomic analysis identifies differentially expressed proteins and reveals potential mechanisms of traumatic heterotopic ossification progression

doi: 10.1016/j.jot.2022.04.003

Figure Lengend Snippet: Representative IHC staining of (a–e) UBA1, EIF3E, RPL27, RPS28, RPL17 in tendon tissue of control group and injury tissue from AT and PAT groups 2days, 7days, 14days, 28days, 10weeks after modeling. Original magnification is 20x. Inserts are approximately 4x magnified images of the boxed area. Scale bars: 250 ​μm.

Article Snippet: Then, the slides were incubated with antibodies against UBA1(1:100, ab180125, Abcam Inc, Cambridge, UK), EIF3E (1:50, ab134958, Abcam Inc, Cambridge, UK), RPL27 (1:50, 14980-1-AP, Proteintech, Chicago, USA), RPL17 (1:300, 67223-1-Ig, Proteintech, Chicago, USA), RPS28(1:50, 14796-1-AP, Proteintech, Chicago, USA) at 4 °C overnight.

Techniques: Immunohistochemistry, Control

Journal: Journal of Orthopaedic Translation

Article Title: Comparative proteomic analysis identifies differentially expressed proteins and reveals potential mechanisms of traumatic heterotopic ossification progression

doi: 10.1016/j.jot.2022.04.003

Figure Lengend Snippet: Representative IHC staining of (a–e) UBA1, EIF3E, RPL27, RPS28, RPL17 in ligament tissue of clinically relevant HO patients, clinically irrelevant HO patients and HO negative patients. Semiquantitative analysis of immunohistochemical staining of UBA1, EIF3E, RPL27, RPS28, RPL17 expression in tissue samples of control, AT and PAT rat models (f) and clinical specimens (g). ImageJ software was applied to calculate the average optical density. Original magnification is 20x. Inserts are approximately 4x magnified images of the boxed area. Scale bars: 250 ​μm n ​= ​3/group. ∗ represent significant change in the expression level compared to control group, ∗P ​< ​0.05, ∗∗P ​< ​0.01.

Article Snippet: Then, the slides were incubated with antibodies against UBA1(1:100, ab180125, Abcam Inc, Cambridge, UK), EIF3E (1:50, ab134958, Abcam Inc, Cambridge, UK), RPL27 (1:50, 14980-1-AP, Proteintech, Chicago, USA), RPL17 (1:300, 67223-1-Ig, Proteintech, Chicago, USA), RPS28(1:50, 14796-1-AP, Proteintech, Chicago, USA) at 4 °C overnight.

Techniques: Immunohistochemistry, Immunohistochemical staining, Staining, Expressing, Control, Software