Attenuated liver fibrosis in the absence of B cells
J. Clin. Invest. Tatiana I. Novobrantseva, et al. 115:3072 doi:10.1172/JCI24798 [Go to this article.]

Figure 4
Collagen production by T6-HSC can be stimulated by B cell–derived soluble factors. T6-HSC cells were seeded at 25% confluence in a 96-well plate in RPMI, 5% FBS, penicillin, streptomycin. After 24 hours, an equal volume of B cell–conditioned medium was added with [³H]-thymidine (to measure proliferation) or with [³H]-proline (to measure de novo collagen synthesis). After 18 hours, the amount of incorporated ³H was quantified. Collagen synthesis per cell is expressed in arbitrary units; we took the ratio of radioactive counts of [³H]-proline/radioactive counts of [³H]-thymidine, considering this ratio equal to 1 in the absence of TGF-β. Each treatment was performed in triplicate. A representative experiment out of 3 is shown. (A) Human TGF-β that is recognized by rat TGF-β receptor was added at different concentration to the culture of T6-HSC cells. [³H]-thymidine incorporation is shown in red. Collagen synthesis as assessed by the rate of [³H]-proline incorporation is shown in blue. T6-HSC proliferation (B) and collagen synthesis relative to cell number as assessed by the ratio of [³H]-proline/[³H]-thymidine incorporation (C) is measured in medium alone (M), with LPS, and with supernatant of B cells cultured with LPS (S). B cells were cultured for 3.5 days prior to harvesting supernatant. Red and blue bars show a mean from triplicate values; error bars show standard deviation.