Immunostaining was performed on cultured BMDMs and frozen sections of human and mouse tissues fixed by 2% paraformaldehyde following a previously described protocol.[18 ,
22 (link)
] In brief, sections were labeled overnight with various combinations of directly conjugated primary antibodies as follows: fluorescein isothiocyanate (FITC)‐conjugated anti‐CD68 antibody (sc‐20060 FITC, Santa‐cruz), FITC‐conjugated anti‐F4/80 (11‐4801‐81, eBioscience), Cy3‐conjugated α‐SMA antibody (C6198, Sigma), Runx1 (sc‐365644, Santa Cruz), p‐Runx1(Ser397) (PA5‐105609, Invitrogen) detected with APC (A21240, Invitrogen) or PE‐conjugated secondary antibodies (A11003, Invitrogen). Sections were washed and, in some cases, DNA was counterstained with DAPI and observed under a fluorescence microscope (Carl Zeiss Axio Observer Z1) or confocal microscope (Carl Zeiss LSM 880).
22 (link)
] In brief, sections were labeled overnight with various combinations of directly conjugated primary antibodies as follows: fluorescein isothiocyanate (FITC)‐conjugated anti‐CD68 antibody (sc‐20060 FITC, Santa‐cruz), FITC‐conjugated anti‐F4/80 (11‐4801‐81, eBioscience), Cy3‐conjugated α‐SMA antibody (C6198, Sigma), Runx1 (sc‐365644, Santa Cruz), p‐Runx1(Ser397) (PA5‐105609, Invitrogen) detected with APC (A21240, Invitrogen) or PE‐conjugated secondary antibodies (A11003, Invitrogen). Sections were washed and, in some cases, DNA was counterstained with DAPI and observed under a fluorescence microscope (Carl Zeiss Axio Observer Z1) or confocal microscope (Carl Zeiss LSM 880).
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