Colony formation of PV ECFCs expressing different forms of PTP-MEG2
. | . | No. of GFP-positive colonies . | . | . | . | |||
|---|---|---|---|---|---|---|---|---|
| ECFCs . | Cytokine(s) . | Small . | Medium . | Large . | Total . | |||
| GFP | SCF | 4 ± 2 | 0 | 0 | 4 ± 2 | |||
| GFP | EPO | 32 ± 5 | 5 ± 2 | 0 | 37 ± 6 | |||
| GFP | SCF+EPO | 39 ± 6 | 45 ± 8 | 14 ± 6 | 98 ± 12 | |||
| MEG2 | SCF | 7 ± 3 | 0 | 0 | 7 ± 3 | |||
| MEG2 | EPO | 34 ± 7 | 3 ± 2 | 0 | 37 ± 8 | |||
| MEG2 | SCF+EPO | 41 ± 9 | 51 ± 10 | 14 ± 4 | 106 ± 15 | |||
| MEG2(D→A) | SCF | 5 ± 2 | 0 | 0 | 5 ± 2 | |||
| MEG2(D→A) | EPO | 20 ± 3 | 0* | 0 | 20 ± 3* | |||
| MEG2(D→A) | SCF+EPO | 46 ± 8 | 32 ± 5* | 2 ± 2* | 80 ± 9* | |||
| MEG2(C→S) | SCF | 2 ± 1 | 0 | 0 | 2 ± 1 | |||
| MEG2(C→S) | EPO | 16 ± 4 | 1 ± 1* | 0 | 17 ± 4* | |||
| MEG2(C→S) | SCF+EPO | 36 ± 7 | 26 ± 4* | 1 ± 1* | 63 ± 9* | |||
. | . | No. of GFP-positive colonies . | . | . | . | |||
|---|---|---|---|---|---|---|---|---|
| ECFCs . | Cytokine(s) . | Small . | Medium . | Large . | Total . | |||
| GFP | SCF | 4 ± 2 | 0 | 0 | 4 ± 2 | |||
| GFP | EPO | 32 ± 5 | 5 ± 2 | 0 | 37 ± 6 | |||
| GFP | SCF+EPO | 39 ± 6 | 45 ± 8 | 14 ± 6 | 98 ± 12 | |||
| MEG2 | SCF | 7 ± 3 | 0 | 0 | 7 ± 3 | |||
| MEG2 | EPO | 34 ± 7 | 3 ± 2 | 0 | 37 ± 8 | |||
| MEG2 | SCF+EPO | 41 ± 9 | 51 ± 10 | 14 ± 4 | 106 ± 15 | |||
| MEG2(D→A) | SCF | 5 ± 2 | 0 | 0 | 5 ± 2 | |||
| MEG2(D→A) | EPO | 20 ± 3 | 0* | 0 | 20 ± 3* | |||
| MEG2(D→A) | SCF+EPO | 46 ± 8 | 32 ± 5* | 2 ± 2* | 80 ± 9* | |||
| MEG2(C→S) | SCF | 2 ± 1 | 0 | 0 | 2 ± 1 | |||
| MEG2(C→S) | EPO | 16 ± 4 | 1 ± 1* | 0 | 17 ± 4* | |||
| MEG2(C→S) | SCF+EPO | 36 ± 7 | 26 ± 4* | 1 ± 1* | 63 ± 9* | |||
Five hundred day-5 PV ECFCs infected with recombinant retroviruses carrying GFP control, PTP-MEG2, PTP-MEG2(D→A), and PTP-MEG2(C→S) were cultured in methylcellulose colony culture medium in the presence of SCF (100 ng/mL), EPO (4 units/mL), or SCF plus EPO. Erythroid cell colonies were counted 9 days later in situ using an inverted fluorescence microscope. Colonies were divided into small (50-100 cells), medium (100-500 cells), and large (>500 cells) categories. Data represent mean ± SD of at least 4 independent experiments. *Significantly different from the correspondent control (GFP and MEG2) data (P < .05).