Fig. 1.

Pre-OCs are more susceptible to TXNRD1 inhibitors than BMDMs, and safe doses of TXNRD1 inhibitors for pre-OCs do not impair osteoclast differentiation and bone resorption.

a, schema for osteoclast differentiation stages in vitro. Analysis was conducted at D0 (BMDMs), D3 (pre-OCs), and D5 (ma-OCs). (b to c), the viability of BMDMs and pre-OCs was measured after 24 h of incubation with AF (b) or TRi-1 (c). n = 3 per group. d, PI staining of BMDMs and pre-OCs was measured after 24 h of incubation with AF (1.0 μM, 2.0 μM) or TRi-1 (2.0 μM, 5.0 μM). n = 3 per group. (e to g), BMDMs incubated with mediums containing M-CSF, RANKL, and maximum safe doses of AF or TRi-1 for pre-OCs. TRAP staining and F-actin formation assay of ma-OCs on day 5 with quantification of TRAP-positive cell area (%) (f) and osteoclast size (mm²) (g). Scale bar, 50 μm. n = 3 per group. (h to i), bone resorption assay was conducted on bovine cortical bone slices and the bone slices were scanned on day 7 (h). Scale bar, 50 μm. n = 3 per group. Bone resorption area (%) (i) were used to evaluate the effects of different dose of AF and TRi-1 on bone resorption capacity. All data in this figure are represented as mean ± SD. ns, no significance, **P < 0.01, ***P < 0.001, ****P < 0.0001.