Figure 3. B. abortus induces HIF‐1α stabilisation and BNIP3L expression in HeLa cells.

• A
  Representative confocal micrographs of HeLa cells infected or not with B. abortus 544 GFP (red) for 24, 48 and 72 h then fixed and immunostained for HIF‐1α (Alexa 568—green). DNA was stained with Hoechst 33258 (Blue). Scale bars: 20 μm.
• B
  Quantification of the percentages of cells positive for a nuclear localisation of HIF‐1α from HeLa cells infected or not (NI) with B. abortus 544 GFP for 24, 48 and 72 h from micrographs shown in (A). Data are presented as means ± SD from n = 3 (biological replicates) independent experiments (the numbers indicated in the columns represent the number of cells analysed per condition). Statistical analyses were performed using a two‐way ANOVA followed by a Šidàk's multiple comparisons test; asterisks indicate significant differences compared to the control (NI); ****P < 0.0001.
• C
  Representative confocal micrographs of HeLa cells infected or not with B. abortus 544 GFP (red) for 48 h, then fixed and immunostained for BNIP3L (Alexa 568—green) and TOMM20 (Alexa 633—magenta). DNA was stained with Hoechst 33258 (blue). Scale bars: 20 μm.
• D
  Relative median fluorescence intensity (MFI) of BNIP3L immunostaining from HeLa cells infected or not (NI) with B. abortus 544 GFP for 48 h as measured by flow cytometry. Data are presented as means ± SD from n = 4 (biological replicates) independent experiments (8,296 cells analysed in total per condition). Statistical analyses were performed using a one sample t‐test; *P < 0.05 (P = 0.0351).

Source data are available online for this figure.