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Impact of Salubrinal on Cell Mortality and Gene Expression in Osteoblasts

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This study investigates the influence of Salubrinal at various concentrations on cell mortality ratios and the relative abundance of osteocalcin (OCN) mRNA in osteoblast-like cells. Findings reveal the effects of Salubrinal on apoptosis pathways, including p-eIF2α and ATF4 expression levels. The experiments utilize Alizarin red S staining and evaluate changes in relative cell numbers, highlighting the therapeutic potential of Salubrinal in osteoblast survival and differentiation. Detailed results are illustrated in multiple figures, emphasizing dose-dependent responses.

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Impact of Salubrinal on Cell Mortality and Gene Expression in Osteoblasts

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  1. a b 2.5 7 6 2.0 5 1.5 4 cell mortality ratio (%) relative cell number 3 1.0 2 0.5 1 0 0 CN 0.1 0.5 1 5 10 20 30 Sal (mM) CN 0.1 0.5 1 5 10 20 30 Sal (mM) c 8 ** 6 Alizarin red S staining per total area (%) ** 4 2 ** ** 0 control 0.1 0.5 1 5 10 20 mM Salubrinal Fig. S1

  2. a control Sal 1 2 4 8 24 1 2 4 8 24 h p-eIF2a Sal 8 h (clone 14) eIF2a ATF4 b-actin b Relative mRNA abundance OCN 2.0 ATF4 2.0 ** ** ** 1.5 * ** 1.5 ** 1.0 1.0 0.5 0.5 0 CN Sal5 Sal10 Sal20 0 CN Sal5 Sal10 Sal20 Fig. S2

  3. 4 * a b ** 3 cell mortality ratio (%) 2 2.5 1 2.0 0 1.5 CN 5 10 20 relative cell number Gu (mM) 1.0 0.5 6.0 OCN ** 0 c CN 5 10 20 5.0 Gu control Gu (mM) 4.0 Relative mRNA abundance 2 4 8 24 2 4 8 24 h 3.0 p-eIF2a 2.0 * 1.0 eIF2a 0 CN 5 10 20 Gu (mM) ATF4 b-actin d Fig. S3

  4. a 7 b 6 5 cell mortality ratio (%) 4 3 2 1 0 CN Gu Sal RANKL c CN Sal Gu RANKL NFATc1 2.0 b-actin 1.5 relative cell number d 1.0 0.5 0 Relative mRNA abundance Gu Sal CN RANKL NFATc1 TRAP 10 300 200 5 100 0 0 CN Sal RANKL Gu CN Sal RANKL Gu Fig. S4

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