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. 2023 Feb 4;14:616. doi: 10.1038/s41467-023-36168-x

Fig. 4. Recombinant La promotes osteoclast fusion.

Fig. 4

(a) Representative fluorescence images of human osteoclasts 3 days post RANKL addition without or with the overnight (end of day 2 post RANKL) addition of recombinant heat-inactivated La 1-408, La 1-408, La 1-375 or La 1-375 Q20A/Y24A/D33I. Recombinant proteins were added at ~40 nM at the end day 2 post-RANKL addition, and cells were fixed the next morning. (Magenta = Phalloidin-Alexa488, Grey = Hoechst) (b) Quantification of a. (inactivated n = 2; La 1-408 n = 4, others n = 3,) (P = 0.1232, 0.0015, 0.0035 and 0.0491, respectively) (c) Quantification of the fraction of nuclei in fused cells that were present in syncytia of various sizes from a. (n = 3). (d) Quantification of the number of fusion events with or without the addition of La 1-187 or La 188-375. Recombinant proteins were added at ~40 nM at the end day 2 post-RANKL addition, and cells were fixed the next morning. (n = 4) (P = 0.36 and 0.0002) (e) The quantification of synchronized fusion events (as illustrated in Fig. 3d) without (wash) and with addition of recombinant La species. “LPC” – indicates that the hemifusion inhibitor was left until fixation. (LPC and La 1−375 n = 3; Wash and La 1-408 n = 4) (P = 0.001 and 0.03, respectively.) (b–e) Statistical significance was evaluated via one-tailed paired t-test. In (b, d, e) the data were normalized to those in control (no protein added in b, d, and wash with no proteins added e). Data are presented as mean values + /- SEM. Source data are provided as a Source Data file.