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. 2017 Jun 21;8:15831. doi: 10.1038/ncomms15831

Figure 1. Plexin-B1 mediates CIL between osteoclasts and osteoblastic cells.

Figure 1

(a) Cartoon representation of culture inserts used for co-culture of osteoclasts with primary osteoblasts or MC3T3-E1 cells. (b) Phase-contrast images acquired immediately and 12 h after lifting the culture insert showing migration of MC3T3-E1 cells towards osteoclasts. Scale bar, 50 μm. (c) Comparison of Plexin-B1 expression in wild-type (WT), Cas9, KO1 and KO2 MC3T3-E1 cells. Tubulin was used as a loading control. (d,e) Phase-contrast images showing a MC3T3-E1 WT (d) or MC3T3-E1 KO1 (e) cell in contact with an osteoclast (OC). Insets are magnified to show changes in cell morphology and migration between time of contact and 20, 30 and 40 min after contact. Scale bar, 50 μm. White triangles point to the site of cell–cell contact. (f) Percentage of WT, Cas9, KO1 and KO2 MC3T3-E1 cells that show collapse of protrusions within 30 min after contact with an osteoclast. n=209–301. (g) Separation between MC3T3-E1 cells and osteoclasts 40 min after contact. n=41–45. (h) Cartoon describing Cx. (i) Cx values for WT cells, Cas9, KO1 and KO2 MC3T3-E1 cells without (WT only) and after contact with osteoclasts. n=20. For g,i, means±s.e.m. are shown. ***P<0.001 and *P<0.05, NS, not significant, Student’s t-test.