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. 2023 Jul 6;75:101773. doi: 10.1016/j.molmet.2023.101773

Figure 2.

Figure 2

Intracellular fatty acids induce HILPDA in adipocytes via ER stress. HILPDA protein level in SVF-derived adipocytes treated with A) 10 μM isoproterenol and B) forskolin for 3 h. C) Hilpda mRNA level after treatments. HILPDA protein level in SVF-derived adipocytes treated for 3 h with 10 μM isoproterenol in the presence and absence of D) 10 μM actinomycin D or E) 10 μM AH7614. HILPDA protein level in SVF-derived adipocytes treated with F) 10 μM isoproterenol for 3 h or G) 10 μM forskolin for 2 h, in the presence or absence of 50 μM Atglistatin (ATGLi). H) NEFA levels in medium of SVF-derived adipocytes after treatment with 10 μM isoproterenol, in the presence or absence of 50 μM Atglistatin (ATGLi) or 20 μM T863 (DGAT1i) for 3 h. N = 3. I) HILPDA protein level in SVF-derived adipocytes treated with 20 μM T863 (DGAT1i) and 10 μM PF-06424439 (DGAT2i) for 10 h in the presence or absence of 50 μM Atglistatin. J) HILPDA protein level in SVF-derived adipocytes treated with 50 μM Atglistatin, 20 μM T863 (DGAT1i), or 10 μM PF-06424439 (DGAT2i) for 10 h. K) Hilpda mRNA expression in SVF-derived adipocytes treated with DGAT1/DGAT2 inhibitors for 10 h. N = 3. HILPDA protein level in SVF-derived adipocytes treated with DGAT1/DGAT2 inhibitors for 10 h in the presence or absence of L) 10 μM etomoxir or 1 μM 10,12-tricosadiynoic acid, or M) 10 μM leupeptin or MG132. N) mRNA expression of Hilpda, Ddit3, Xbp1s and Atf4 in 3T3-L1 adipocytes treated with DGAT1/DGAT2 inhibitors for different durations. N = 3. HILPDA protein level in 3T3-L1 adipocytes treated with O) DGAT1/DGAT2 inhibitors for 10 h, P) 10 μM isoproterenol for 3 h, and Q) 5 μM thapsigargin for 12 h, in the presence or absence of specific ER stress pathway inhibitors. Western blots were probed with antibodies against HILPDA and HSP90. Asterisk indicates significantly different from control treatment according to Student's t-test. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.