Figure 3: Knock-out of NM1 causes increased resistance to hypotonic conditions.

(A) WT and NM1 KO mice were used for preparation of mouse skin fibroblasts. By exogenous expression of NM1-Flag in WT and NM1 KO cell lines, WT+NM1 and NM1 KO+NM1, respectively were prepared by lentivirus transduction. NM1 KO+GFP was used as a negative control (B). All cell lines were subjected to hypotonic conditions for 1 min followed by 2 hours of recovery under normal growth conditions. NM1 KO fibroblasts show significantly higher ability to resist hypotonic conditions than WT cells. Moreover, WT fibroblasts expressing extra NM1-Flag have lower ability to survive than WT cells. NM1 KO fibroblasts expressing NM1-Flag show similar survival rate as WT, which correlates with the amount of NM1 expressed by both cell lines. The KO-GFP cell line was used as a negative control of the lentivirus transduction effect on hypotonic stress survival. All measurements were done in triplicates. Values are shown as mean ± S.D and statistical significance is presented as ^ns p > 0.05, **p < 0.01, ****p < 0.0001, respectively.