Extended Data Fig. 3: LIFR deficiency or overexpression does not affect hepatocyte proliferation ex vivo or in vitro.
a-e. Primary hepatocytes isolated from Lifrfl/fl and Lifrfl/fl;Alb-Cre mice were cultured for 3 hours, followed by treatment with 100 ng/mL of Hgf and/or 20 ng/mL of Egf for 48 hours. a-c. qPCR of mRNA of Lifr (a), cyclin D1 (b), and Pcna (c) in Hgf- and/or Egf-treated hepatocytes. n = 4, 4, 5, 5, 5, 5, 4, and 4 biological replicates. d, e. Immunofluorescence staining of Ki67 (d; overlay with DAPI staining) and percentage of Ki67-positive cells (e) in Hgf- and/or Egf-treated hepatocytes. Scale bars, 100 μm. n = 4 biological replicates. f-j. Primary hepatocytes isolated from C57BL/6J mice 10 days after injection with control adenovirus or LIFR-expressing adenovirus were cultured for 3 hours, followed by treatment with 100 ng/mL of Hgf and/or 20 ng/mL of Egf for 48 hours. f-h. qPCR of mRNA of Lifr (f), cyclin D1 (g), and Pcna (h) in Hgf- and/or Egf-treated hepatocytes. n = 5, 5, 4, 4, 4, 4, 5, and 5 biological replicates. i, j. Immunofluorescence staining of Ki67 (i; overlay with DAPI staining) and percentage of Ki67-positive cells (j) in Hgf- and/or Egf-treated hepatocytes. Scale bars, 100 μm. n = 5 biological replicates. k-m. qPCR of mRNA of Lifr (k), cyclin D1 (l), and Pcna (m) in Hgf- and/or Egf-treated hepatocytes isolated from C57BL/6J mice. The cells were infected with control adenovirus or LIFR-expressing adenovirus for 24 hours before Hgf and/or Egf treatment. n = 5, 5, 4, 4, 4, 4, 5, and 5 biological replicates. Statistical significance in a-c, e-h, and j-m was determined by a two-tailed unpaired t-test. Error bars are s.e.m.
