Fig. 5: UBIAD1 interacted with the C-terminus of H-Ras.

a FNTA knockdown decreased interaction between UBIAD1 and H-Ras. HEK293T cells were transfected with 3Flag-UBIAD1 and GFP-H-Ras with or without sh-FNTA. Seventy-two hours after transfection, the cell lysate was exposed to antibodies and analyzed by IP and IB as indicated. The same experiment was repeated three times. b Icmt knockdown decreased interaction between UBIAD1 and H-Ras. HEK293T cells were transfected with 3Flag-UBIAD1 and GFP-H-Ras with or without si-Icmt. Seventy-two hours after transfection, the cell lysate was exposed to antibodies and analyzed by IP and IB as indicated. The same experiment was repeated three times. c 2-BP treatment increased interaction between UBIAD1 and H-Ras. HEK293T cells were transfected with 3Flag-UBIAD1 and GFP-H-Ras, followed by 72 h of culture and 24 h of culturing with or without 2BP. The cell lysate was exposed to antibodies and analyzed by IP and IB as indicated. The same experiment was repeated three times. d Interaction of UBIAD1 with truncated H-Ras. HEK293T cells were transiently transfected with plasmids as indicated. Forty-eight hours after transfection, the cell lysate was exposed to antibodies and analyzed by IP and IB as indicated. The same experiment was repeated three times. e The C-terminus of H-Ras decreased interaction between UBIAD1 and H-Ras. HEK293T cells were transiently transfected with plasmids as indicated. Forty-eight hours after transfection, the cell lysate was exposed to antibodies and analyzed by IP and IB as indicated. The same experiment was repeated three times. f The C-terminus of H-Ras blocked UBIAD1 function with decreases in p-ERK levels. HEK293T or T24 cells were transfected with plasmids as indicated. Twenty-four hours after transfection, the total cell lysate was analyzed by WB. The lower panel presents the ratio (mean ± SD) from densitometry analyses. **p < 0.01, ns: not significant, Student’s t-test, n = 3 experiments. g The C-terminus of H-Ras suppressed the UBIAD1-induced decrease in T24 cell viability. T24 cells were transfected with plasmids as indicated. Cell viability was detected by the MTT assay. **p < 0.01, Student’s t-test, n = 3 experiments. h The C-terminus of H-Ras inhibited the UBIAD1-induced decrease in T24 cell proliferation. T24 cells were transfected with the plasmids as indicated. Cell proliferation was detected using the BrdU cell proliferation ELISA kit. **p < 0.01, Student’s t-test, n = 3 experiments. i The C-terminus of H-Ras inhibited the UBIAD1-induced decrease in T24 cell growth. T24 cells were transfected with plasmids as indicated. Cell growth was detected by cell counting, n = 3 experiments