American Society of Hematology

Fig. 4.

$Fig. 4. Relative amounts of hyaluronan in cell layers from healthy donor and multiple myeloma patient–derived bmMPC cultures. / FACE analysis of HA in the cell layer ethanol precipitate fractions of bmMPC cultures derived from a healthy donor (lanes 2 and 3) and a myeloma patient (lanes 4 and 5). Cultures were incubated as indicated in the presence (+; lanes 3 and 5) or absence (−; lanes 2 and 4) of dexamethasone (DEX) as described in “Materials and methods.” All samples were digested with hyaluronidase SD, chondroitinase ABC, and glucoamylase as described in “Materials and methods.” No AMAC-derivatized bands were detected in the absence of enzyme digestion (data not shown), similar to the results shown in Figure 3. The gel image in panel A was overexposed to show minor bands, whereas the gel image in panel B was exposed such that all pixels were within the linear 12-bit depth range required for quantitation. In panel B, the AMAC-derivatized glucose bands in lanes 4 and 5 were covered to allow imaging of the AMAC-derivatized ΔDiHA bands. The percent HA detected as AMAC-derivatized ΔDiHA in each culture is listed below each lane in panel B and is normalized to the (−) DEX myeloma value. Note the presence of a prominent unknown band in the samples from the multiple myeloma patient (X1). AMAC-derivatized standards (lanes 1 and 6) include, from top to bottom,N-acetylgalactosamine, maltotetraose, maltotriose, maltose, glucose, ΔDiHA, ΔDi0S, 6-sulfatedN-acetylgalactosamine, and 4-sulfatedN-acetylgalactosamine. Each lane represents either 2.5% (healthy donor) or 5% (myeloma patients) of the total cell layer samples. Data presented are representative of 4 healthy donors and 4 myeloma patients. Similar results were observed in all donors.$

Relative amounts of hyaluronan in cell layers from healthy donor and multiple myeloma patient–derived bmMPC cultures.

FACE analysis of HA in the cell layer ethanol precipitate fractions of bmMPC cultures derived from a healthy donor (lanes 2 and 3) and a myeloma patient (lanes 4 and 5). Cultures were incubated as indicated in the presence (+; lanes 3 and 5) or absence (−; lanes 2 and 4) of dexamethasone (DEX) as described in “Materials and methods.” All samples were digested with hyaluronidase SD, chondroitinase ABC, and glucoamylase as described in “Materials and methods.” No AMAC-derivatized bands were detected in the absence of enzyme digestion (data not shown), similar to the results shown in Figure 3. The gel image in panel A was overexposed to show minor bands, whereas the gel image in panel B was exposed such that all pixels were within the linear 12-bit depth range required for quantitation. In panel B, the AMAC-derivatized glucose bands in lanes 4 and 5 were covered to allow imaging of the AMAC-derivatized ΔDiHA bands. The percent HA detected as AMAC-derivatized ΔDiHA in each culture is listed below each lane in panel B and is normalized to the (−) DEX myeloma value. Note the presence of a prominent unknown band in the samples from the multiple myeloma patient (X1). AMAC-derivatized standards (lanes 1 and 6) include, from top to bottom,N-acetylgalactosamine, maltotetraose, maltotriose, maltose, glucose, ΔDiHA, ΔDi0S, 6-sulfatedN-acetylgalactosamine, and 4-sulfatedN-acetylgalactosamine. Each lane represents either 2.5% (healthy donor) or 5% (myeloma patients) of the total cell layer samples. Data presented are representative of 4 healthy donors and 4 myeloma patients. Similar results were observed in all donors.

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