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To examine whether treating cartilage tissue with collagenase modifies cathepsin activity, we incubated purified recombinant human cathepsins B and S separately with collagenase media, as previously done with IC/DC samples.
Authenticated immortalized human mammary epithelial (HMLE) cells expressing empty vector (pWZL), Snail, Snail shControl, or Snail shFOXC2 cells were cultured in MEGM media as previously described (Werden et al., 2016).
We cultured the cells on STO-feeders in a PGC-defined media as previously described [50].
The HL-1 cardiomyocyte line (generous gift of W. Claycomb) was maintained in Claycomb media as previously described [16].
HL-1 cardiomyocytes were grown in 5% CO2 at 37°C in supplemented Claycomb media as previously described [41].
For co-culture experiments, the cell lines were grown to confluence in 6-well tissue culture plates in their respective growth media as previously described.
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These cells were differentiated into mature adipocytes and conditioned media were collected as previously described (see Methods).
Sediments were inoculated on Loewenstein-Jensen (LJ) media with and without pyruvate and on Middlebrook 7H11 media supplemented as previously described [ 20].
All media used were either YPD (high phosphate) or phosphate-depleted YPD media, made as previously described [ 48].
Zebrafish (AB) were mated and grown at 28.5°C in embryo rearing media (E3) as previously described in [18].
The different SMC phenotypes were induced via changes in media components, as previously demonstrated elsewhere.
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