Journal: Cell Stress & Chaperones

Article Title: Heat shock-induced PI(4)P increase drives HSPA1A translocation to the plasma membrane in cancer and stressed cells through PI4KIII alpha activation

doi: 10.1016/j.cstres.2025.100130

Figure Lengend Snippet: PI4KIII alpha transcript and protein levels remain stable post-heat shock. (a) qPCR analysis of PI4KIII alpha transcript stability using separate batches of cells from all other experiments, including RNA-seq. Transcript levels were assessed at control (37 °C), 0 h post-heat shock, and 8 h recovery and normalized to housekeeping genes (ACTB and GAPDH). HSPA1A was included as a positive control for heat shock response. Statistical significance was determined using one-way ANOVA (Analysis of Variance) followed by the post-hoc Tukey HSD (Honestly Significant Difference) test. (b) Representative Western blot of total PI4KIII alpha protein levels in cell lysates under control conditions, with 0 h post-heat shock and 8 h recovery. HSPA1A is a positive control, demonstrating heat shock-induced upregulation, while GAPDH verifies equal protein loading (complete blots in ). (c) Quantification of Western blot results from independent experiments showing relative PI4KIII alpha and HSPA1A protein levels normalized to GAPDH. Closed circles represent individual data points. Center lines indicate medians; box limits represent the 25th and 75th percentiles; whiskers extend 1.5 times the interquartile range; crosses denote sample means. Statistical significance was determined using one-way ANOVA (Analysis of Variance) followed by the post-hoc Tukey HSD (Honestly Significant Difference) test (N.S, not significant).

Article Snippet: • Anti-GAPDH Monoclonal Antibody (Mouse IgG2b; 60004-1-Ig; dilution 1:1000), Proteintech Group, Inc (Rosemont, IL, USA) Blots were incubated with primary antibodies overnight (∼16 h) at 4 °C with constant rotation.

Techniques: RNA Sequencing, Control, Positive Control, Western Blot