Figure 1.
Highly expressed ZDHHC21 specifically regulates OXPHOS hyperactivity in AML cells. (A) The ATP level of healthy donors and patients with AML. (B) Schematic representation of the compound screen. Cellular ATP levels were measured by an enhanced ATP Assay Kit. ATP levels were normalized to their protein concentrations. (C) Relative OXPHOS inhibition in AML cells and HSCs. HL60 cells and HSCs were treated with a collection of 267 PTM inhibitors (10 μM) for 24 hours. Compounds that selectively inhibited OXPHOS in AML cells rather than HSCs are indicated with red circles. Rotenone and IACS-010759, indicated with yellow solid circles, were the positive controls to suppress OXPHOS. (D) The inhibitory effect of 2BP on ATP production in AML cells. HL60 cells were treated with 2BP at a series of concentrations (0, 10, 20, 40, 60 μM), and other cells lines were treated at a single concentration, including NB4 (60 μM), MV411 (40 μM), OCI-AML3 (60 μM), THP-1 (60 μM), and Kasumi-1 (60 μM). ATP levels were normalized to their protein concentrations. (E) Mitochondrial OCR of HL60 cells after treatment with control (DMSO) or 2BP for 24 hours. (F) Relative OXPHOS inhibition in AML cells and HSCs. HL60 cells and HSCs were transfected with small interfering RNAs targeting ZDHHCs for 72 hours. (G) Top: the protein levels of ZDHHC21 were measured by western blotting to evaluate the silencing efficiency of different shRNA (#1 and #2) against ZDHHC21 in HL60 cells for 5 days. Bottom: cellular ATP levels of shZDHHC21-transfected HL60 cells for 5 days. ATP levels were normalized to their protein concentrations. (H) Relative fold change of gene expression for 173 patients with AML and 70 normal controls obtained from the GEPIA database. The phylogenetic tree represents the sequential evolutionary relationships of a set of ZDHHC family. (I) The expression levels of ZDHHC1, ZDHHC21, and ZDHHC23 were analyzed in 31 cancer species using the GEPIA database. (J) Mitochondrial OCR of HL60 cells infected with lentivirus-shZDHCH21s for 5 days. Data are presented as mean ± SD (n = 3). n.s., P > .05; ∗P < .05; ∗∗P < .01; ∗∗∗P < .001. The significance analysis was conducted by a two-tailed unpaired t test or one-way analysis of variance. AA, antimycin A; Ctrl, control; FCCP, fluoro-carbonyl cyanide phenylhydrazone; rot, rotenone.

Highly expressed ZDHHC21 specifically regulates OXPHOS hyperactivity in AML cells. (A) The ATP level of healthy donors and patients with AML. (B) Schematic representation of the compound screen. Cellular ATP levels were measured by an enhanced ATP Assay Kit. ATP levels were normalized to their protein concentrations. (C) Relative OXPHOS inhibition in AML cells and HSCs. HL60 cells and HSCs were treated with a collection of 267 PTM inhibitors (10 μM) for 24 hours. Compounds that selectively inhibited OXPHOS in AML cells rather than HSCs are indicated with red circles. Rotenone and IACS-010759, indicated with yellow solid circles, were the positive controls to suppress OXPHOS. (D) The inhibitory effect of 2BP on ATP production in AML cells. HL60 cells were treated with 2BP at a series of concentrations (0, 10, 20, 40, 60 μM), and other cells lines were treated at a single concentration, including NB4 (60 μM), MV411 (40 μM), OCI-AML3 (60 μM), THP-1 (60 μM), and Kasumi-1 (60 μM). ATP levels were normalized to their protein concentrations. (E) Mitochondrial OCR of HL60 cells after treatment with control (DMSO) or 2BP for 24 hours. (F) Relative OXPHOS inhibition in AML cells and HSCs. HL60 cells and HSCs were transfected with small interfering RNAs targeting ZDHHCs for 72 hours. (G) Top: the protein levels of ZDHHC21 were measured by western blotting to evaluate the silencing efficiency of different shRNA (#1 and #2) against ZDHHC21 in HL60 cells for 5 days. Bottom: cellular ATP levels of shZDHHC21-transfected HL60 cells for 5 days. ATP levels were normalized to their protein concentrations. (H) Relative fold change of gene expression for 173 patients with AML and 70 normal controls obtained from the GEPIA database. The phylogenetic tree represents the sequential evolutionary relationships of a set of ZDHHC family. (I) The expression levels of ZDHHC1, ZDHHC21, and ZDHHC23 were analyzed in 31 cancer species using the GEPIA database. (J) Mitochondrial OCR of HL60 cells infected with lentivirus-shZDHCH21s for 5 days. Data are presented as mean ± SD (n = 3). n.s., P > .05; ∗P < .05; ∗∗P < .01; ∗∗∗P < .001. The significance analysis was conducted by a two-tailed unpaired t test or one-way analysis of variance. AA, antimycin A; Ctrl, control; FCCP, fluoro-carbonyl cyanide phenylhydrazone; rot, rotenone.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal