Extended Data Fig. 2: UBXN9 does not affect myotube gene expression or regulate RLR signaling in macrophages and human lung A549 cells.
From: UBXN9 governs GLUT4-mediated spatial confinement of RIG-I-like receptors and signaling
a, Diagram illustrating the design of Ubxn9flox/flox mice. b, Immunoblots of UBXN9 and housekeeping tubulin proteins in various tissues of Ubxn9−/− mice ( + TMX) compared to Ubxn9+/+ littermates ( + corn oil). c, Protocol for isolation of primary mouse myoblast from Ubxn9+/+ and Ubxn9−/− mice. d, Representative light microscopic images demonstrating pre-plating steps described in (c). Scale bar, 10μm. Cells were isolated from Ubxn9+/+ mice as an example of the purification process. DM, differentiation medium. e, Immunoblots of UBXN9 and GAPDH in purified myoblasts isolated from Ubxn9+/+ and Ubxn9−/− mice. f, Myf5 and MyoG mRNA expression in Ubxn9+/+ and Ubxn9−/− myoblasts and myotubes, respectively (n = 3 Ubxn9+/+ and 4 Ubxn9−/− mice). g, Immunoblots of UBXN9 and β-actin in bone marrow-derived macrophages (BMDMs) isolated and differentiated from Ubxn9+/+ and Ubxn9−/− mice (n = 2 mice/group as a representative of UBXN9 knockout efficiency). h, Immunoblots of UBXN9 and β-actin in UBXN9+/+ and UBXN9−/− A549 lung cells. i, Cellular Ifnb1 transcript levels in Ubxn9+/+ and Ubxn9−/− BMDMs (n = 3 mice/group) before and after transfection with poly(I:C) (left) or 3p-hpRNA (right). j, IFN-β protein levels from UBXN9+/+ and UBXN9−/− A549 cells (n = 2 biological replicates from two independent experiments) stimulated with poly(I:C) for 12 h. The data are representative of one (f) and two independent experiments (g-j). ns, not significant based on two-way ANOVA with Šidák’s multiple comparisons test (i). Bar: mean ± SEM.
