-
A, B
Proliferation ability of GSTZ1 knockout (GSTZ1‐KO) HepG2 cells treated with or without different doses of picropodophyllin (PPP, 0.5 μM and 1.0 μM) for 24 h. (A) Cell growth curves. (B) Representative images (left) and quantification (right) were obtained by EdU incorporation assays. Scale bars: 50 μm.
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C
Flow cytometry detection of apoptosis, conducted using Annexin V–FITC/PI double staining and quantification, shows early apoptotic cells in GSTZ1‐OE Huh7 (top) and GSTZ1‐KO HepG2 (bottom) cells. GSTZ1‐KO cells were treated with PPP as described above.
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D, E
Heat map of RNA‐sequencing data (described in Fig
4) (D) and quantitative RT–PCR (E) show the differential expression of downstream signaling molecules and apoptosis‐associated genes in IGF1R‐mediated antiapoptotic pathway. The asterisks indicate statistically significant differences between AdGSTZ1 and AdGFP.
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F, G
Expression of IGF1R downstream signaling molecules (F) and apoptosis‐associated proteins (G) in GSTZ1‐OE Huh7 (left) and GSTZ1‐KO HepG2 (middle, right) cells. GSTZ1‐KO cells were treated with or without PPP (1.0 μM) for 24 h (middle) and transfected with control (shCon) or IGF1R shRNA (shIGF1R) (right). All cells were treated with (F) or without (G) IGF‐1 (Beyotime, 50 ng/ml) for 1 h after serum starvation overnight.
Data information: Values are shown as means ± SD (
n = 3), *
P < 0.05, **
P < 0.01, ***
P < 0.001, Student's
t‐test (two groups) or one‐way ANOVA followed by the Tukey test (more than two groups). Abbreviations: DAPI, 4′,6‐diamidino‐2‐phenylindole; EdU, 5‐ethynyl‐2′‐deoxyuridine; ns, not significant; p‐IGF1Rβ, phospho (Tyr1131)‐IGF1Rβ; p‐AKT, phospho (Thr308)‐AKT; p‐ERK1/2, phospho (Thr202/Tyr204)‐ERK 1/2; p‐BAD, phospho (Ser136)‐BAD; p‐CREB, phospho (Ser133)‐CREB; BCL2, B cell lymphoma/leukemia‐2 protein; CASP, caspase.
Source data are available online for this figure.
