Extended Data Fig. 8: Inhibition of p38, ROCK1, or Gβγ ameliorates lactate-induced browning in adipocytes.
From: Activation of GPR81 by lactate drives tumour-induced cachexia
a, Representative western blots and statistic results of UCP1 and activation of p38-ATF2 in the SVF-derived adipocytes after treatment with 20 mM sodium L-lactate, NaCl, or sodium D-lactate (n = 6 biologically independent samples in each group). b, Representative western blots and ratio of phosphorylated to total ATF2 in the SVF-derived adipocytes, showing that lactate-induced activation of p38-ATF2 was not blocked by PKA inhibitor H89 (n = 4 biologically independent samples in each group). c, Tissue weights of inguinal white adipose tissue (iWAT), gonad white adipose tissue (gWAT), and quadriceps of mice implanted with osmotic pumps loaded with PBS or sodium L-lactate and treated with H89 (n = 6 for each group). d-g, Statistic results showing that in the SVF-derived adipocytes, treatment with inhibitor for Gαi (PTX) or Gβγ (Gallein) (d, n = 5 biologically independent samples in each group), silencing the expression of Gβ1 or Gβ2 by siRNA (e, n = 5 biologically independent samples in each group), silencing the expression of RhoA or ROCK1 by specific siRNA (f, n = 5 biologically independent samples in each group), or treatment with inhibitor for p38 (SB203580) or ROCK1 (Y-27632) (g, n = 5 biologically independent samples in each group) abolished the lactate-induced activation of ATF2. h,i, Immunofluorescence staining (h) and statistic result of the signal intensity (i) of colocalized RhoA and Rhotekin in the SVF-derived adipocytes (n = 6 biologically independent samples in each group). Scale bars, 20 μm. j, Weights of LLC tumours derived from mice injected in the inguinal fat pad with inhibitor for p38, ROCK1, or Gβγ (n = 6 for each group). All data are presented as the mean ± s.e.m. P values were determined by one-way ANOVA with Tukey’s multiple comparisons (a-g, i, j).
