Skip to main content
. 2019 Feb 26;41:538–555. doi: 10.1016/j.ebiom.2019.02.009

Fig. 1.

Fig. 1

Highly expressed PLEKHO1 in osteoblasts of RA patients and CIA mice. (a,b) Comparison of PLEKHO1 levels in bone tissues from knee joint between RA and TM patients underwent knee joint replacement surgery by western blot. a, Representative electrophoretic bands of the samples from fifteen RA patients and eight TM patients are shown. The numbers below the bands represent a semiquantitative value. b, The semiquantitative data of the protein levels. (c,d) Comparison of PLEKHO1 expression (red) within osteocalcin positive (Ocn+) osteoblasts (green) in bone tissues from knee joint between RA and TM patients by immunofluorescence analysis. c, Representative fluorescent micrographs of the samples from five RA patients and five TM patients are shown. Merged images with DAPI staining showed co-staining of PLEKHO1 and Ocn+ osteoblasts (arrows, yellow). H&E staining of the same sections is shown, and black arrowheads point to bone-formation surfaces, enriched by those cells with pink, which is a merged colour of green (osteoblast marker) and blue (DAPI staining for nuclei) in the immunofluorescence staining. Scale bars, 20 μm. d, Comparison of the percentage of cells co-expressing Ocn and PLEKHO1 in Ocn+ cells from bone tissues of knee joint between RA patients and TM patients. (e,f) Comparison of PLEKHO1 expression (red) within Ocn+ osteoblasts (green) in bone tissues of ankle joint from hind paws of CIA and normal mice on day 49 after primary immunization by immunofluorescence analysis. e, Representative fluorescent micrographs of the bone samples from ten CIA mice and ten normal mice are shown. Merged images with DAPI staining showed co-staining of PLEKHO1 and Ocn+ osteoblasts (arrows, yellow). H&E staining of the same sections is shown, and red arrowheads point to bone-formation surfaces. Scale bars: 20 μm. f, Comparison of the percentage of cells co-expressing Ocn and PLEKHO1 in Ocn+ cells from bone tissues of ankle joint from hind paws between CIA mice and normal mice on day 49 after primary immunization. (g) Time course changes of Plekho1 mRNA level in Ocn+ osteoblasts from CIA and normal mice by laser-capture microdissection (LCM) combined with qPCR. (h) Levels of Plekho1 mRNA induced by proinflammatory cytokines (IL-1β, IL-6 and TNF-α) in human osteoblast–like cell line MG-63 (left) and mouse osteoblast–like cell line MC3T3E1 (right) by real time PCR analysis. N: no cytokines, served as control. Note: RA, rheumatoid arthritis; TM, severe trauma. All data are the mean ± s.d. *P < 0.05, **P < 0.01. Two-way ANOVA with Bonferroni posttests was performed. Comparisons between two groups were performed using a Student's t-test. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)