![HA-induced cluster formation is dependent on the expression of long-stretch CD44 variants. (A) Flow cytometric analysis of the expression of CD44 and its variants on MM cell lines. The means ± SD (bars) of 3 independent experiments are shown. (B) MM cells were cultured at 1 × 105 cells/mL in the presence of 120 ng/mL of HA under shear stress for 1 hour. Left panel: Representative phase-contrast images of MM cells. Bar represents 200 μm. Right panel: The means ± SD (bars) of the size of the cell clusters (n = 8-20). (C) KMM.1 sublines were cultured at 1 × 105 cells/mL with the indicated concentrations of HA under shear stress for 1 hour. Left panel: Representative phase-contrast images of 4 KMM.1 sublines cultured with 120 ng/mL of HA. Bar represents 400 μm. Right panel: The means ± SD (bars) of the size of cell clusters (n = 8-20). ∗P < .05 by one-way ANOVA with Tukey’s multiple comparison test. (D) Flow cytometric analysis of the expression of CD44 and its variants on primary MM cells isolated from the BM (except for EMD-MNC, which was isolated from the EMD lesion) of patients with (+) or without (−) EMD. In a patient with UPN5, BM samples were isolated at diagnosis [EMD (−)] and before the development of EMD [EMD (+)] (Supplemental Figure 2C). ∗P < .05 by one-way ANOVA with Tukey’s multiple comparison test between EMD (+) and EMD (−) groups.](https://ash.silverchair-cdn.com/ash/content_public/journal/bloodadvances/7/4/10.1182_bloodadvances.2022007291/7/blooda_adv-2022-007291-gr4.jpeg?Expires=1735843768&Signature=wGgsjChe69gTPI4Jlp7rnFLjoc1h1APhj2~hpYPEDvnWUBwJ5LX6s7wJKYYOKViQkKycsgC00TPqByjh04tbYse5-Uq0Y3PIe-Ge0AvsUr5Q5dE8aEN6EHlc5MYR80ii2g1VdBoOLPwWk9R47QGdpvXHTwuPUqWw9eT9l1c8FTk1SjUYiqLIupq-1lAQtPC0m8B9Kc9OxR2kMYd5RwytAN8gyBufPFcoK90Jp4Ro3K5dut6gBa0L~A9Za~vFX-ItB63NHPrRHq8GCGNRqFR5GPnFoZ6pV9RmC5hAP5mFnRLQ9ZuSBgwiPP~mMjodKXXHmy8Ir2YdfnAvkasY14tp3w__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
HA-induced cluster formation is dependent on the expression of long-stretch CD44 variants. (A) Flow cytometric analysis of the expression of CD44 and its variants on MM cell lines. The means ± SD (bars) of 3 independent experiments are shown. (B) MM cells were cultured at 1 × 105 cells/mL in the presence of 120 ng/mL of HA under shear stress for 1 hour. Left panel: Representative phase-contrast images of MM cells. Bar represents 200 μm. Right panel: The means ± SD (bars) of the size of the cell clusters (n = 8-20). (C) KMM.1 sublines were cultured at 1 × 105 cells/mL with the indicated concentrations of HA under shear stress for 1 hour. Left panel: Representative phase-contrast images of 4 KMM.1 sublines cultured with 120 ng/mL of HA. Bar represents 400 μm. Right panel: The means ± SD (bars) of the size of cell clusters (n = 8-20). ∗P < .05 by one-way ANOVA with Tukey’s multiple comparison test. (D) Flow cytometric analysis of the expression of CD44 and its variants on primary MM cells isolated from the BM (except for EMD-MNC, which was isolated from the EMD lesion) of patients with (+) or without (−) EMD. In a patient with UPN5, BM samples were isolated at diagnosis [EMD (−)] and before the development of EMD [EMD (+)] (Supplemental Figure 2C). ∗P < .05 by one-way ANOVA with Tukey’s multiple comparison test between EMD (+) and EMD (−) groups.
