Expression of SHH and Hh signaling pathway components in MM cells. (A) Flow cytometry analysis for identification and selection of CD138^–CD19^–, CD138^–CD19⁺, and CD138⁺CD19^– cell populations from MM patients’ BM aspirates for expression of SHH. (B) Percentages of SHH-positive cells in each of the 3 cell populations in BM samples from 6 more MM patients. (C) RT-PCR showing the levels of mRNA for Hh signaling components in highly purified CD138⁺ primary MM cells from 10 MM patients (Pt MM) and 6 MM cell lines (MMCLs). (D) Real-time PCR showing the levels of mRNA for Hh signaling components in patients’ highly purified CD19⁺ B cells (Pt B cell; n = 4) or CD138⁺ primary MM cells (Pt MM; n = 10) from MM patients’ BM aspirates, and 6 MMCLs. (E) Western blotting showing protein expression of SHH, PTCH, GLI1, in MMCLs and purified CD19⁺ B cells from healthy donor. GAPDH, glyceraldehyde phosphate dehydrogenase.