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. Author manuscript; available in PMC: 2017 Sep 15.
Published in final edited form as: J Immunol. 2016 Aug 17;197(6):2522–2531. doi: 10.4049/jimmunol.1600915

Figure 2. HS1 activation, F-actin polymerization and receptor internalization assays.

Figure 2

(A) Representative histograms of HS1-Y397 phosphorylation (P-HS1) flow cytometry analysis at 2 and 5 minutes following anti-IgM or anti-IgD receptor stimulation in one U-CLL case. Isotypes are shown, as control. (B) Mean ± SEM of P-HS1, after 2 and 5 minutes of anti-IgM or anti-IgD stimulation, as analyzed by flow cytometry on 7 U-CLL cases, and expressed as percentage (%) of the untreated control. *P <0.05; Mann-Whitney test. (C) Western Blot analysis of P-HS1 at the activatory tyrosine residue Y397 and HS1 total levels after 2 and 5 minutes of anti-IgM or anti-IgD stimulation. Two representative U-CLL cases (U-CLL#1, U-CLL#2) out of 5 tested are shown. (D) Mean ± SEM of F-actin polymerization after a period of 5 minutes of anti-IgM or anti-IgD stimulation, as analyzed on 4 U-CLL cases, and expressed as % of the untreated control. *P <0.05; unpaired t test. (E) Representative histograms of IgM and IgD surface staining of one U-CLL case before and after 5 minutes of anti-IgM, anti-IgD or anti-Igκ stimulation. Isotypes are shown, as control. (F) Comparison of the effect of heavy chain stimulation (anti-IgM, anti-IgD) and light chain stimulation (anti-Igκ) on surface IgM and IgD levels. Mean ± SEM of IgM and IgD receptor expression, expressed as Mean Fluorescent Intensity Ratio (MFIR) before and after 5 minutes of anti-IgM, anti-IgD or anti-Igκ, as analyzed in 4 Igκ+ U-CLL cases.