[Retracted] TRPM7 Upregulate the Activity of SMAD1 through PLC Signaling Way to Promote Osteogenesis of hBMSCs

Figure 4

The expression of osteogenic markers and the osteogenic potential of hBMSCs under inhibiting TRPM7 and activating PLC (HSPG2). (a) (A) Western blot analysis of PLC activity in cells induced with osteogenic medium or osteogenic medium plus control shRNA or TRPM7 shRNA induced for 14 days. (B) The relative activity of PLC protein was quantified and plotted. (b) (A) Western blot analysis of the activity of PLC in cells induced with osteogenic medium or osteogenic medium plus Naltriben or 2-APB induced for 14 days. (B) The relative activity of PLC protein was quantified and plotted. Control shRNA: hBMSCs transfected with negative control-shRNA-vector; TRPM7 shRNA: hBMSCs transfected with TRPM7-shRNA-vector. (c) RT-PCR analysis of osteogenic markers ALP, COL1A1, RUNX2, OPN, and OCN in cells induced with osteogenic medium supplemented with different pharmaceuticals induced for 14 days. (d) (A) Western blot analysis of osteogenic marker proteins ALP, COL1A1, and RUNX2 in cells induced with osteogenic medium supplemented with different pharmaceuticals induced for 14 days. (B) The relative levels of osteogenic marker proteins were quantified and plotted. (e) Analysis of ALP staining in cells induced with osteogenic medium supplemented with different pharmaceuticals induced for 14 days. (f) Analysis of ALP activity (the unit for p-nitrophenyl: μmol) in cells induced with osteogenic medium supplemented with different pharmaceuticals induced for 14 days. (g) Staining of ARS in cells induced with osteogenic medium supplemented with different pharmaceuticals induced for 14 days. (h) A quantification of ARS in cells induced with osteogenic medium supplemented with different pharmaceuticals induced for 14 days. Scale: 200 μm. Control: cells induced in osteogenic medium without any pharmacological treatment; M-3M3FBS: cells induced in osteogenic medium supplemented with 20 μM M-3M3FBS; 2-APB+M-3M3FBS: cells induced in osteogenic medium supplemented with 50 μM 2-APB and 20 μM/mL M-3M3FBS; 2-APB: cells induced in osteogenic medium supplemented with 50 μM 2-APB. , , and .

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)