Effects of lentivirally encoded shRNAs on colony formation of CML CD34+ cells and normal CD34+ cells
. | RFP+ colonies/total colony number, % . | . | . | |
---|---|---|---|---|
Sample, dcH1-shRNA-SR . | High cytokine concentration . | Low cytokine concentration . | Comparison of RFP positivity between low and high cytokine condition, % . | |
CML CD34+ cells | ||||
A (b3a2) | ||||
GL4 | 33 | 33 | 100 | |
b3a2 | 21 | 13 | 62 | |
237SHP2 | 30 | 12 | 40 | |
B (b3a2) | ||||
GL4 | 23 | 21 | 92 | |
b3a2 | 17 | 9 | 53 | |
237SHP2 | 34 | 17 | 50 | |
C (b3a2) | ||||
GL4 | 25 | 21 | 84 | |
b3a2 | 21 | 10 | 48 | |
594SHP2 | 56 | 20 | 36 | |
714SHP2 | 11 | 5 | 45 | |
D (b3a2) | ||||
GL4 | 44 | 38 | 86 | |
b3a2 | 33 | 16 | 48 | |
714SHP2 | 22 | 11 | 50 | |
E (b2a2) | ||||
GL4 | 24 | 20 | 84 | |
GFP | 22 | 19 | 86 | |
594SHP2 | 15 | 5 | 33 | |
STAT5 | 16 | 6 | 37 | |
E.2 (b2a2) | ||||
GL4 | 56 | 51 | 91 | |
237SHP2 | 70 | 40 | 57 | |
Gab2 | 49 | 27 | 55 | |
STAT5 | 31 | 16 | 52 | |
F (b2a2) | ||||
GL4 | 11 | 10 | 91 | |
GFP | 11 | 10 | 91 | |
237SHP2 | 5 | 2 | 40 | |
594SHP2 | 3 | 0 | 0 | |
STAT5 | 7 | 0 | 0 | |
G (b2a2) | ||||
GL4 | 38 | 36 | 95 | |
b3a2 | 46 | 38 | 83 | |
237SHP2 | 14 | 2 | 14 | |
STAT5 | 13 | 4 | 31 | |
H (b3a2) | ||||
GL4 | 57 | 50 | 88 | |
b3a2 | 50 | 23 | 46 | |
237SHP2 | 43 | 23 | 53 | |
594SHP2 | 58 | 27 | 46 | |
Gab2 | 25 | 18 | 72 | |
STAT5 | 64 | 10 | 16 | |
I (b3a2) | ||||
GL4 | 32 | 33 | 103 | |
b3a2 | 6 | 0 | 0 | |
237SHP2 | 15 | 0 | 0 | |
J (b2a2) | ||||
GL4 | 50 | 54 | 108 | |
237SHP2 | 23 | 15 | 65 | |
Gab2 | 24 | 12 | 50 | |
STAT5 | 19 | 9 | 47 | |
K (b2a2) | ||||
GL4 | 12 | 11 | 92 | |
237SHP2 | 63 | 32 | 51 | |
Gab2 | 24 | 12 | 50 | |
STAT5 | 33 | 14 | 42 | |
L (b3a2) | ||||
GL4 | 34 | 33 | 96 | |
b3a2 | 11 | 3 | 27 | |
237SHP2 | 11 | 6 | 54 | |
Normal CD34+ cells | ||||
A | ||||
GL4 | 47 | 40 | 85 | |
b3a2 | 34 | 30 | 88 | |
237SHP2 | 28 | 26 | 93 | |
714SHP2 | 27 | 22 | 82 | |
B | ||||
GL4 | 29 | 25 | 86 | |
237SHP2 | 9 | 10 | 111 | |
594SHP2 | 12 | 13 | 108 | |
714SHP2 | 16 | 15 | 94 | |
Gab2 | 25 | 23 | 92 | |
C | ||||
GL4 | 53 | 52 | 98 | |
b3a2 | 30 | 35 | 117 | |
237SHP2 | 27 | 25 | 93 | |
D | ||||
GL4 | 34 | 35 | 103 | |
237SHP2 | 21 | 21 | 100 | |
594SHP2 | 21 | 23 | 109 | |
Gab2 | 20 | 17 | 85 | |
STAT5 | 26 | 19 | 73 | |
E | ||||
GL4 | 32 | 35 | 109 | |
594SHP2 | 28 | 32 | 114 | |
STAT5 | 42 | 32 | 76 | |
F | ||||
GL4 | 28 | 27 | 96 | |
Gab2 | 25 | 25 | 100 | |
STAT5 | 16 | 14 | 88 |
. | RFP+ colonies/total colony number, % . | . | . | |
---|---|---|---|---|
Sample, dcH1-shRNA-SR . | High cytokine concentration . | Low cytokine concentration . | Comparison of RFP positivity between low and high cytokine condition, % . | |
CML CD34+ cells | ||||
A (b3a2) | ||||
GL4 | 33 | 33 | 100 | |
b3a2 | 21 | 13 | 62 | |
237SHP2 | 30 | 12 | 40 | |
B (b3a2) | ||||
GL4 | 23 | 21 | 92 | |
b3a2 | 17 | 9 | 53 | |
237SHP2 | 34 | 17 | 50 | |
C (b3a2) | ||||
GL4 | 25 | 21 | 84 | |
b3a2 | 21 | 10 | 48 | |
594SHP2 | 56 | 20 | 36 | |
714SHP2 | 11 | 5 | 45 | |
D (b3a2) | ||||
GL4 | 44 | 38 | 86 | |
b3a2 | 33 | 16 | 48 | |
714SHP2 | 22 | 11 | 50 | |
E (b2a2) | ||||
GL4 | 24 | 20 | 84 | |
GFP | 22 | 19 | 86 | |
594SHP2 | 15 | 5 | 33 | |
STAT5 | 16 | 6 | 37 | |
E.2 (b2a2) | ||||
GL4 | 56 | 51 | 91 | |
237SHP2 | 70 | 40 | 57 | |
Gab2 | 49 | 27 | 55 | |
STAT5 | 31 | 16 | 52 | |
F (b2a2) | ||||
GL4 | 11 | 10 | 91 | |
GFP | 11 | 10 | 91 | |
237SHP2 | 5 | 2 | 40 | |
594SHP2 | 3 | 0 | 0 | |
STAT5 | 7 | 0 | 0 | |
G (b2a2) | ||||
GL4 | 38 | 36 | 95 | |
b3a2 | 46 | 38 | 83 | |
237SHP2 | 14 | 2 | 14 | |
STAT5 | 13 | 4 | 31 | |
H (b3a2) | ||||
GL4 | 57 | 50 | 88 | |
b3a2 | 50 | 23 | 46 | |
237SHP2 | 43 | 23 | 53 | |
594SHP2 | 58 | 27 | 46 | |
Gab2 | 25 | 18 | 72 | |
STAT5 | 64 | 10 | 16 | |
I (b3a2) | ||||
GL4 | 32 | 33 | 103 | |
b3a2 | 6 | 0 | 0 | |
237SHP2 | 15 | 0 | 0 | |
J (b2a2) | ||||
GL4 | 50 | 54 | 108 | |
237SHP2 | 23 | 15 | 65 | |
Gab2 | 24 | 12 | 50 | |
STAT5 | 19 | 9 | 47 | |
K (b2a2) | ||||
GL4 | 12 | 11 | 92 | |
237SHP2 | 63 | 32 | 51 | |
Gab2 | 24 | 12 | 50 | |
STAT5 | 33 | 14 | 42 | |
L (b3a2) | ||||
GL4 | 34 | 33 | 96 | |
b3a2 | 11 | 3 | 27 | |
237SHP2 | 11 | 6 | 54 | |
Normal CD34+ cells | ||||
A | ||||
GL4 | 47 | 40 | 85 | |
b3a2 | 34 | 30 | 88 | |
237SHP2 | 28 | 26 | 93 | |
714SHP2 | 27 | 22 | 82 | |
B | ||||
GL4 | 29 | 25 | 86 | |
237SHP2 | 9 | 10 | 111 | |
594SHP2 | 12 | 13 | 108 | |
714SHP2 | 16 | 15 | 94 | |
Gab2 | 25 | 23 | 92 | |
C | ||||
GL4 | 53 | 52 | 98 | |
b3a2 | 30 | 35 | 117 | |
237SHP2 | 27 | 25 | 93 | |
D | ||||
GL4 | 34 | 35 | 103 | |
237SHP2 | 21 | 21 | 100 | |
594SHP2 | 21 | 23 | 109 | |
Gab2 | 20 | 17 | 85 | |
STAT5 | 26 | 19 | 73 | |
E | ||||
GL4 | 32 | 35 | 109 | |
594SHP2 | 28 | 32 | 114 | |
STAT5 | 42 | 32 | 76 | |
F | ||||
GL4 | 28 | 27 | 96 | |
Gab2 | 25 | 25 | 100 | |
STAT5 | 16 | 14 | 88 |
Cultures were stimulated with high (20 ng/mL GM-CSF, 10 ng/mL IL-3) or low cytokine concentration (0.2 ng/mL GM-CSF, 0.1 ng/mL IL-3). The right column indicates the relative amount of the RFP+ colony ratio under low stimulation (ratio at high concentration set as 100% for each lane).