Figure 1.

Mild hypothermia of 35°C promoted the neuronal differentiation of cultured hNSCs

(A and B) Representative immunofluorescence images showing β-tubulin III, DCX, S100β, and GFAP positive differentiated hNSCs at Day 14 with 37°C-, 35°C-, and 33°C-treatments.

(C–F) The percentage of β-tubulin III, DCX, S100β, and GFAP positive cells in (A) and (B).

(G–J) qRT-PCR analysis showing the relative mRNA expression of TUBB3, DCX, S100B, and GFAP of cultured hNSCs at Day 14 with 37°C-, 35°C-, and 33°C-treatments.

(K) Western blots showing the protein expression of β-tubulin III, DCX, and GFAP of cultured hNSCs at Day 14 with 37°C-, 35°C-, and 33°C-treatments. β-actin was used as the loading control.

(L) The quantification of western blots of (K). Normalized β-tubulin III, DCX, and GFAP to corresponding loading control were summarized for three independent trials.

(M) Representative immunofluorescence images showing β-tubulin III and DCX positive cells from Day 0 to Day 14 every two days with 35°C-treatment.

(N–Q) The percentage of β-tubulin III, DCX, GFAP, and SOX2 positive cells from Day 0 to Day 14 every two days with 35°C- and 37°C-treatment. All data presented as mean ± SD. One-way ANOVA tests were used in (C–J) and (L). Two-way ANOVA tests were used in (N–Q). NS, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Scale bars represent 50 μm in (A), (B), and (M).

See also Figures S1 and S2.