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Fig. 4 | BMC Cancer

Fig. 4

From: Identification of markers that functionally define a quiescent multiple myeloma cell sub-population surviving bortezomib treatment

Fig. 4

a Western blots for GRP78 protein in RPMI8226 cells. Due to abundance of GRP78 in RPMI8226 cells, 2 μg of protein per well (instead of 20 μg in other blots) was used in all blots. b IF Detection of GRP78 in Bz-surviving RPMI8226 cells. Fluorescence intensity quantification is shown as fold increase over the negative control. c Western blots for GRP78 protein in U266 cells 6 days after Bz washout. d Detection and quantification (right graph) of GRP78 in H2B-GFPHIGH label-retaining cells at 3 and 6 days after drug washout by IF. **** p < 0.0001 (unpaired t test). Scale bar =25 μm. e Detection of GRP78 in cytospins from bone marrow aspirates of MM patients. Representatives of each group (low, medium, and high GRP78 levels) are shown here. Scale bar =20 μm. f Graphical representation of patient groups via GRP78 MFI per cell. Symbols represent stage of each patient. Patient MM# numbers are shown adjacent to each symbol. P < 0.0001 between groups (one-way ANOVA). g Western blots showing depletion of GRP78 protein in RPMI8226 cells after treatment with SubAB toxin. Non-functional mutant SubAA272B was used as a control. GAPDH and β-Actin were used as loading controls. h Cell viability plot of Bz-pulsed RPMI8226 cells, +/- GRP78 depletion via treatment (at two different concentrations) with SubAB toxin. Non-functional mutant SubAA272B was used as a control. Trypan blue exclusion was used as viability assay

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