Fig. 1. Osteoclastogenic differentiation is accompanied by drastic changes in the steady-state levels and localization of La molecular species.

a Representative images of stages of osteoclastogenic derivation of human monocytes after M-CSF (6 days, referred to as “M-CSF”) and after M-CSF (6 days) followed by M-CSF + RANKL (5 days, “RANKL”), respectively. (Magenta = Phalloidin-Alexa488, Cyan = Hoechst). b Quantification of the number of fusion events normalized to the total number of nuclei observed over time following RANKL addition. (n = 3) Each point represents an average of >7500 nuclei scored. c Representative Bis-Tris PAGE separation and silver staining of whole protein lysates from M-CSF derived osteoclast precursors and at 3 days post RANKL application. Lysates were ran until the 50 KDa marker nearly ran off the 4–12% polyacrylamide gel to achieve maximal separation of proteins at this molecular weight, leading to the band of interest appearing misleadingly heavy. <denotes band of interest excised from both lanes and evaluated via mass spectrometry. d Representative Tris-Glycine Western blot with α-La mAb evaluating La expression in whole protein lysates from primary human monocytes and the osteoclastogenic stages depicted in a. e Representative Tris-Glycine Western blot with α-La mAb evaluating the time course of La expression following RANKL addition. (α-GAPDH loading control). f Representative immunofluorescence images of La in M-CSF derived osteoclast precursors and at 3 days post RANKL application (α-La mAb). g Representative immunofluorescence images of La in forming osteoclasts 2–5 days post RANKL application (α-La mAb). Cells were stained for La at the described timepoints with membrane permeabilization. Source data are provided as a Source Data file.