Figure S3.
PI3KC2α is required for the growth of BRAF-mutant melanoma cells. (A and B) RT–qPCR (A) and immunoblot (B) detecting mRNA and protein levels of PI3KC2α after knockdown in A375 and SK-MEL-28 cells, respectively. One-way ANOVA, Bonferroni’s test. (C) Relative cell number of A375 and SK-MEL-28 cells after PI3KC2α knockdown. Cell viability was measured by the CCK8 assay after 72 h (n = 3). One-way ANOVA, Bonferroni’s test. (D) Colony formation assay of A375 and SK-MEL-28 cells with PI3KC2α knockdown. Colonies were stained with crystal violet after 14 days. (E) Immunofluorescence of PI(3,4)P2 after PI3KC2α knockdown in A375 cells (n = 3). Scale bar, 20 μm. One-way ANOVA, Dunnett’s test. (F) Immunoblot of indicated proteins in A375 cells after PI3KC2α knockdown. (G) Immunoblot of indicated proteins after endogenous PI3KC2α knockdown with or without the overexpression of HA-PI3KC2α WT, S329A, or S329D mutant in A375 and SK-MEL-28 cells. (H) Relative cell number of A375 and SK-MEL-28 cells after endogenous PI3KC2α knockdown with or without the overexpression of HA-PI3KC2α WT, S329A, or S329D mutant. Cell viability was measured by the CCK8 assay after 72 h (n = 3). One-way ANOVA, Bonferroni’s test. (I) Colony formation assay of A375 and SK-MEL-28 cells depleted of endogenous PI3KC2α with or without the overexpression of HA-PI3KC2α WT, S329A, or S329D. Colonies were stained with crystal violet after 14 days. ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05; P > 0.05; not significant (ns). Source data are available for this figure: SourceData FS3. Refer to the image caption for details.

PI3KC2α is required for the growth of BRAF-mutant melanoma cells. (A and B) RT–qPCR (A) and immunoblot (B) detecting mRNA and protein levels of PI3KC2α after knockdown in A375 and SK-MEL-28 cells, respectively. One-way ANOVA, Bonferroni’s test. (C) Relative cell number of A375 and SK-MEL-28 cells after PI3KC2α knockdown. Cell viability was measured by the CCK8 assay after 72 h (n = 3). One-way ANOVA, Bonferroni’s test. (D) Colony formation assay of A375 and SK-MEL-28 cells with PI3KC2α knockdown. Colonies were stained with crystal violet after 14 days. (E) Immunofluorescence of PI(3,4)P2 after PI3KC2α knockdown in A375 cells (n = 3). Scale bar, 20 μm. One-way ANOVA, Dunnett’s test. (F) Immunoblot of indicated proteins in A375 cells after PI3KC2α knockdown. (G) Immunoblot of indicated proteins after endogenous PI3KC2α knockdown with or without the overexpression of HA-PI3KC2α WT, S329A, or S329D mutant in A375 and SK-MEL-28 cells. (H) Relative cell number of A375 and SK-MEL-28 cells after endogenous PI3KC2α knockdown with or without the overexpression of HA-PI3KC2α WT, S329A, or S329D mutant. Cell viability was measured by the CCK8 assay after 72 h (n = 3). One-way ANOVA, Bonferroni’s test. (I) Colony formation assay of A375 and SK-MEL-28 cells depleted of endogenous PI3KC2α with or without the overexpression of HA-PI3KC2α WT, S329A, or S329D. Colonies were stained with crystal violet after 14 days. ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05; P > 0.05; not significant (ns). Source data are available for this figure: SourceData FS3.

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