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. 2012 Aug 10;103(10):1820–1825. doi: 10.1111/j.1349-7006.2012.02374.x

Figure 1.

Figure 1

Prosaposin (PSAP) upregulated estrogen receptor alpha (ERα) expression. (a,b) After the initial 48‐h estrogen deprivation, MCF‐7 and T47D cells were treated with increasing concentrations (0, 5, 20 ng/mL) of PSAP protein with or without E2 for 24 h. Estrogen receptor alpha protein level was detected by western blot. (c,d) MCF‐7 and T47D cells were treated with PSAP (0, 5, 20 ng/mL) for 24 h. The total RNA was extracted and subjected to real‐time polymerase chain reaction (PCR) analysis of ERα mRNA level. (e,f) MCF‐7 cells were transfected with pSilencer 2.1 (pSi) or pSilencer‐PSAP (pSi‐PSAP) plasmid. 48 h after transfection, cell lysates were quantified and blotted with anti‐PSAP, anti‐ERα and anti‐glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) antibodies (e), and the total RNA was extracted and subjected to real‐time PCR analysis of ERα mRNA level (f). (g) MCF7 cells were treated with control or conditioned media (CM) from pcDNA3.1‐PSAP transfected MCF7 cells. Estrogen receptor alpha protein level was detected by western blot. The amount of PSAP in CM was detected by enzyme linked immunosorbent assay (ELISA) assay in (h).