Further characterizing lipid transfer and contact formation by PITPβ. Quantitative data are shown as mean ± SD, with the number of independent experiments indicated. Statistics was performed using the two-tailed Student’s t test: ****P < 0.0001, **P < 0.01, ns (non-significant) P > 0.05. (A) Thrombin-mediated cleavage of the 6x histidine tag from recombinant PITPβ, n = 3. (B) Liposome binding study examining PITPβ incubated with different liposomes, as indicated, n = 3. The pellet (P) fraction contains PITPβ on the membrane, while the supernatant (S) fraction contains soluble PITPβ. (C) Liposome-binding study involving PITPβ forms, as indicated, incubated with Golgi-like liposomes, n = 4. Pellet (P) fraction contains PITPβ on the membrane, while the supernatant (S) fraction contains soluble PITPβ. (D) Reconstitution of lipid transfer and contact formation reveals that PC transfer by PITPβ requires PA incorporated into acceptor (L_b) liposomes, n = 5. (E) Reconstitution of lipid transfer and contact formation revealing that PC transfer by PITPβ requires its FFAT-like motif, n = 3. (F) Reconstitution of lipid transfer and contact formation revealing that PC transfer by PITPβ is not affected by PI(4)P incorporated into acceptor (L_b) liposomes, n = 4. Source data are available for this figure: SourceData FS5.