Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2016 Dec 12;213(13):2967–2988. doi: 10.1084/jem.20151665

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2016 Chan et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

PMC Copyright notice

Figure 2. — MTD chemotherapy–treated CAFs increase the percentage of TICs. (A) BCAF-011 CAFs were treated with MTD-doxorubicin, MTD-paclitaxel, or vehicle as in Fig. 1 B and then co-cultivated with BC-011 or BT-474 carcinoma cells in a dual chamber culture apparatus for 5 d, after which the carcinoma cells were isolated and treated with the same therapy regimen or vehicle for 4 d. The cells surviving the treatment were determined by a CYTOX-orange/Hoechst 33342 two-color fluorescence cell viability assay. Data from two independent experiments (n = 3 in each group) are shown. (B) BCAF-011 CAFs were treated with vehicle (vehicle-CAF), MTD-doxorubicin, MTD-paclitaxel, or MTD–4H-CPA (MTD-CAF) and then co-cultivated with BC-011 carcinoma cells in a dual chamber culture apparatus, and the carcinoma cells were subjected to flow cytometric analysis. (Left) Shown are representative plots showing patterns of CD44 and CD24 staining of carcinoma cells with the frequency of the boxed CD44⁺CD24^−/low cell population as a percentage of cancer cells shown. (Right) The percentages of CD44⁺CD24^−/low cancer cells at different times in the co-culture. Data from two independent experiments (n = 3 in each group) are shown. (C and D) The same co-culture experiments as described in B were performed using different basal (HCC-1954, HCC-1806, and HCC-38 cells; C)- or luminal (BT-474 cells; D)-subtype breast carcinoma cells and BCAF-008 (C and D) or BCAF-011 CAFs (C). Shown are the percentages of CD44⁺CD24^−/low (C) or ALDH^high cancer cells (D) at different times in the co-culture. Data from two independent experiments (n = 3 in each group) are shown. (E) BC-011 carcinoma cells were cultured in the conditioned medium derived from vehicle- or MTD-CAFs in nonadherent culture plates for 10 d. Shown are representative phase contrast images of the resultant tumorspheres. Bars, 100 µm. (F) Limiting dilution assay demonstrating the tumorsphere formation efficiency of BC011 carcinoma cells cultured in the conditioned medium derived from vehicle- or MTD-CAFs. The arrow indicates change of slope of the trend lines, suggestive of differential tumor-sphere formation ability. Data from two independent experiments (n = 6 in each group) are shown. (G) Breast carcinoma cells can be subdivided into four subpopulations according to the expression patterns of CD44, CD24, and ALDH. (H) MTD-CAFs promoted the conversion of nonstem-like carcinoma cells (i.e., non-CD44⁺CD24^−/low/ALDH^low cells in cell subpopulation D) but not the ALDH^high TICs (cell subpopulation B) into CD44⁺CD24^−/low cells. Data from two independent experiments (n = 3 in each group) are shown. (I) Vehicle- or MTD-doxorubicin–treated BCAF-011 or BCAF-008 CAFs were co-inoculated with GFP-transduced BC-011 and BC-008 carcinoma cells, respectively, into the mammary fat pads of NOG mice. 2 wk after cell inoculation, the tumors were removed for cell dissociation, and the cells were subjected to flow cytometric analyses. (J) The percentages of CD44⁺CD24^−/low cells relative to GFP-positive cancer cells in tumors described in I. Data from one experiment (n = 3 in each group) are shown. Data are mean ± SEM; Student’s t test; **, P < 0.01; ***, P < 0.001.