Ab7880

After the removal of the OCT compound, immunofluorescence staining was performed on 20-µm sections using polyclonal rabbit-anti-Ki67 (1:400, ab15580, Abcam), monoclonal anti-mouse-K14 (1:200, ab7880, Abcam), monoclonal anti-rabbit-phospho-STAT3 (pStat3, 1:200, #9145, Cell Signaling Technology), monoclonal anti-mouse-STAT3 (Stat3, 1:200, #9139, Cell Signaling Technology) overnight at 4 °C. Then, Alexa488-conjugated donkey-anti-rabbit IgG (1:400, A21206, Molecular Probes) or Alexa546-conjugated goat-anti-mouse IgG (1:400, A11003, Molecular Probes) was used as a secondary antibody for 3 h at room temperature. The sections were then counterstained with DAPI (1:500, Dojindo) and mounted with fluorescent mounting medium (Dako). At least three embryos of each genotype were used for each analysis.

Immunofluorescence staining was performed on 20-µm sections using polyclonal rabbit-anti-Ki67 (1:400, ab15580, Abcam), monoclonal rabbit anti-K17 (1:200, #4543, Cell Signaling Technology), monoclonal anti-K14 (1:200, ab7880, Abcam), monoclonal rabbit anti-phospho-Stat3 (pStat3, 1:200, #9145, Cell Signaling Technology), monoclonal rabbit anti-Stat3 (1:200, #9139, Cell Signaling Technology) overnight at 4 °C. Then, Alexa488-conjugated goat-anti-rabbit IgG (1:400, A21206, Molecular Probes) or Alexa546-conjugated goat-anti-mouse IgG (1:400, A11003, Molecular Probes) was used as secondary antibody. The sections were then counterstained with DAPI (1:500, Dojindo) and mounted with fluorescent mounting medium (Dako). At least three embryos of each genotype were used for each analysis.

Immunofluorescence staining was performed using polyclonal rabbit-anti-Ki67 (1:400, ab15580, Abcam), polyclonal rabbit anti-K6 (1:200, #4543, 905701, Biolegend), monoclonal anti-K14 (1:200, ab7880, Abcam), monoclonal rabbit anti-phospho-Stat3 (pStat3, 1:200, #9145, Cell Signaling Technology) or monoclonal rabbit anti-Stat3 (1:200, #9139, Cell Signaling Technology) overnight at 4°C. Alexa488-conjugated goat-anti-rabbit IgG (1:400, A21206, Molecular Probes) or Alexa546-conjugated goat-anti-mouse IgG (1:400, A11003, Molecular Probes) was used as secondary antibody. DAPI (1:500, Dojindo) was used for nuclear staining and the sections were mounted with fluorescence mounting medium (Dako). At least three embryos of each genotype were used for each analysis.
The percentage of proliferating cells at the fusing or contacting epithelium between the primary and the secondary palate was determined by counting Ki67-positive cells and reporting this value as a percentage of the total number of cells, as determined by DAPI staining.