Table 3. Effect of cold stress (33°C) and knockdown of mTOR on RBC2 and NC line SC proliferation¹.

Line	Temperature2	Knockdown3	Sampling time				P-value4
			0 h	24 h	48 h	72 h
RBC2	38	Control	0.11a,x ± 0.01	0.17c,x ± 0.01	0.41b,y ± 0.03	0.93c,z ± 0.04	< 0.0001
		mTOR	0.11a,y ± 0.01	0.15d,y ± 0.01	0.22c,y ± 0.03	0.63d,z ± 0.05	< 0.0001
	33	Control	0.11a,w ± 0.01	0.13d,x ± 0.01	0.15cd,y ± 0.15	0.21ef,z ± 0.05	< 0.0001
		mTOR	0.10a,zy ± 0.01	0.09e,y ± 0.01	0.11d,zy ± 0.11	0.12f,z ± 0.05	< 0.0001
NC	38	Control	0.11a,w ± 0.01	0.29a,x ± 0.01	0.55a,y ± 0.03	1.82a,z ± 0.05	< 0.0001
		mTOR	0.11a,w ± 0.01	0.21b,x ± 0.01	0.35b,y ± 0.03	1.10b,z ± 0.05	< 0.0001
	33	Control	0.11a,x ± 0.01	0.18c,y ± 0.01	0.21c,y ± 0.02	0.32e,z ± 0.05	< 0.0001
		mTOR	0.10a,x ± 0.01	0.16cd,y ± 0.01	0.19c,y ± 0.02	0.29e,z ± 0.05	< 0.0001
P-value5		L	0.5281	<0.0001	<0.0001	<0.0001
		T	0.1860	<0.0001	<0.0001	<0.0001
		K	0.9005	<0.0001	<0.0001	<0.0001
		L × T × K	0.9862	0.0143	0.6462	0.0013

¹ Each value represents mean DNA concentration (μg / well) ± Standard error of mean (SEM)

² Incubation temperature (°C) during proliferation and differentiation

³ Control = transfecting cells with a negative control sequence; mTOR = knockdown of mTOR

⁴ Effect of sampling times within each cell line, temperature, and knockdown of mTOR

⁵ Effect of line (L), temperature (T), and knockdown of mTOR (K), and the interaction effect among line, temperature, and knockdown of mTOR (L × T × K) within each sampling time

^a-f Mean DNA concentration (μg / well ± SEM) within a column (sampling time) without a common letter are significantly different

^w-z Mean DNA concentration (μg / well ± SEM) within a row (temperature and cell line) without a common letter are significantly different

P ≤ 0.05 was considered as significant different