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. 2016 Jun 23;35(17):1853–1867. doi: 10.15252/embj.201593148

Figure 4. INPP5E knockdown influences neither lysosomal integrity nor fusion between endosomes and lysosomes.

Figure 4

  1. N1E‐115 cells treated with siControl or siINPP5Es were cultured in growth medium with 20 μg/ml DQ‐BSA for 12 h and chased for 2 h. Cells were fixed and stained with anti‐LAMP1 antibodies and analyzed by immunofluorescence microscopy. Insets show the boxed areas at high magnification. Scale bar, 10 μm.
  2. Quantitation of signal intensities in (A) showing colocalization of DQ‐BSA with LAMP1 (means ± s.d.; n > 50 cells from three independent experiments). Baf.A1 was used as a control for inhibition of delivery and degradation of DQ‐BSA via the endocytic pathway. **P < 0.01; n.s., non‐significant.
  3. N1E‐115 cells treated with siControl, siINPP5Es, or siCHMP5 were stimulated with 100 ng/ml EGF for the indicated time periods and then analyzed by immunoblot using anti‐EGFR and anti‐α‐tubulin antibodies. Levels of CHMP5 mRNA 72 h after transfection of N1E‐115 cells with siControl or siCHMP5 as analyzed by RT–PCR.
  4. Quantitation of EGFR degradation ratio in (C). Results represent means ± s.d. of three independent experiments. **P < 0.01.

Source data are available online for this figure.