Fig. 5: MS3-6 reduces STAT3 nuclear localization. | Nature Communications

Fig. 5: MS3-6 reduces STAT3 nuclear localization.

From: Selective inhibition of STAT3 signaling using monobodies targeting the coiled-coil and N-terminal domains

Fig. 5

a Representative immunoblot analysis of cellular fractionation experiments from three independent experiments. A549 expressing monobodies upon 48 h of 1 µg ml−1 doxycycline treatment were stimulated with either IL-6 or IL-22 for 15 min. Nuclear (RCC1) and cytosolic (Tubulin) fractions were recovered and probed for total, p-Y705 and p-S727 STAT3. Quantification of three independent experiments normalized to the HA4-Y87A monobody control are shown in b and are plotted as mean ± SD. Significance according to a two-tailed unpaired t-test analysis: *P = 0.0278, **P = 0.0028. c Confocal microscopy images from two independent experiments of HEK293 cells expressing a monobody-GFP fusion and treated with IL-6 to assess nuclear translocation. Scale bar represents 10 µm. The right column shows the outlines used to determine nuclear and cellular compartments indicated by blue and green lines, respectively, using the CellProfiler software. The number of individual cells analyzed (n) is indicated on the figure. Quantification of STAT3 nuclear/cytoplasmic levels from two independent experiments is shown in d. Significance according to a two-tailed unpaired t-test analysis: **P = 0.0065, ****P ≤ 0.0001. The number of analyzed cells is indicated below the graph. Source data are provided as a source data file.

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