Fig. 4: Physiological consequence of RHBDL4 ablation.

a TMED7 expression is increased in BMDMs derived from RHBDL4 knockout (R4 ko) mice relative to cells derived from wt animals. Signal for TLR4 was obtained on a separate WB. Actin is used as a loading control. Right panel, quantification of TMED7 in R4 ko cells relative to control (means ± SEM, n = 4; *p < 0.05, two-sided Student’s t test, p = 0.0383). b Amount of TNFα (left) and IL-6 (right) determined by ELISA in the supernatant of wt and R4 ko BMDMs after LPS (100 ng/ml) treatment for 0, 6 (TNFα p = 0.0154, IL-6 p = 0.0419), 12 (TNFα p = 0.001, IL-6 p = 0.0093), and 24 h (TNFα p = 0.002, IL-6 p = 0.0003). Secretion of both cytokines is increased in cells derived from R4 ko mice compared to cells derived from wt mice (means ± SEM, n = 4 except for time points 0 h in wt animals and 24 h in R4 ko animals for IL-6 and timepoint 0 h for wt animals for TNF\({{{{{\rm{\alpha }}}}}}\) that were n = 3; *p < 0.05, **p < 0.01, ***p < 0.001, two-sided Student’s t test). c Relative expression of TNFα in white blood cells from R4 ko mice is increased compared to cells derived from wt mice 150 min after injection of LPS (35 µg/g of body weight). TNFα expression has been normalized to and expressed as a ratio relative to expression in cells derived from wt mice (means ± SEM, n = 5 mice per condition; **p < 0.01, two-sided Student’s t test, p = 0.0092). d R4 ko mice show a decreased survival upon LPS injection (35 µg/g of body weight) when compared to wt mice (n = 12 mice per condition, p = 0.0054, Log-rank, Mantel-Cox test). e TNF\({{{{{\rm{\alpha }}}}}}\) expression is increased in THP-1 cells upon transfection with siRNA targeting RHBDL4 (siR4) relative to control siRNA (sictrl) after infection with M. tuberculosis for 24 h (means ± SEM, n = 3, *p < 0.05, two-sided Student’s t test, p = 0.0132). f Amount of secreted TNF\({{{{{\rm{\alpha }}}}}}\) determined by ELISA in the supernatant of THP-1 cells is increased in cells transfected with siRNA targeting RHBDL4 (siR4) relative to control siRNA (sictrl) upon infection with M. tuberculosis (+Mtb) for 24 h (means ± SEM, n = 3; *p < 0.05, two-sided Student’s t test, p = 0.0191). Source data are provided as a Source Data file.