Figure S5.
Supplementary data toFigs. 7 and 8. (A) Schematic cartoon of mApple-Lamp1-phLuorin. (B) Western blots show the level of CathepsinD in control or two Tex2-KO clones. (C) Left: representative flow cytometry analysis of either control or Tex2 KO HeLa stained with lysotracker using unstained HeLa cells as a control; right: normalized lysotracker fluorescence intensity of control or Tex2 KO cells as the left panel from four independent assays. One sample t test. Mean ± SD. (D) Representative images of either control (top), two Tex2-KO clones (middle), or Tex2-KO-1 cells treated with BafA1 expressing GFP–LC3–RFP were labeled with Lysotracker (blue) with insets. (E) Pearson’s correlation coefficient of GFP–LC3 vs. Lysotracker in control (10 cells), Tex2 KO-1 (10 cells), Tex2 KO-2 (10 cells), or Tex2 KO-1 treated with BafA1 (10 cells) in three independent assays. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. (F) Representative images of either control (top) or Tex2 KO-1 (bottom) cells expressing GFP-Lact2-C2 (green; PS sensor) with insets. (G) Pearson’s correlation coefficient of GFP-Lact2-C2 vs. Lamp1-mCh (18 control; 20 Tex2-KO cells). Two-tailed unpaired Student’s t test. Mean ± SD. (H) Representative images of either control (top) or Tex2 KO-1 (bottom) cells expressing GFP-P4M (PI4P sensor) with insets. (I) Pearson’s correlation coefficient of GFP-P4M vs. either Lamp1-mCh (13 control; 31 Tex2-KO cells) or B4GalT1-TM-BFP (Golgi marker; 13 control; 21 Tex2-KO cells). Two-tailed unpaired Student’s t test. Mean ± SD. (J) Linescan analysis of GFP-P4M at LE/lys/Golgi relative to the PM in control or Tex2 KO-1 HeLa cells, as shown in H. (K) Representative images of either control (top) or Tex2 KO-1 (bottom) cells expressing PI(4,5)P2 sensor PLCδ-PH-GFP with insets. (L) Linescan analysis of PLCδ-PH-GFP at LE/lys relative to the PM in control or Tex2 KO-1 HeLa cells, as shown in K. Two-tailed unpaired Student’s t test. Mean ± SD. Scale bar, 10 μm in the whole cell images and 2 μm in the insets in D, F, H, and K. Source data are available for this figure: SourceData FS5.

Supplementary data to Figs. 7 and 8. (A) Schematic cartoon of mApple-Lamp1-phLuorin. (B) Western blots show the level of CathepsinD in control or two Tex2-KO clones. (C) Left: representative flow cytometry analysis of either control or Tex2 KO HeLa stained with lysotracker using unstained HeLa cells as a control; right: normalized lysotracker fluorescence intensity of control or Tex2 KO cells as the left panel from four independent assays. One sample t test. Mean ± SD. (D) Representative images of either control (top), two Tex2-KO clones (middle), or Tex2-KO-1 cells treated with BafA1 expressing GFP–LC3–RFP were labeled with Lysotracker (blue) with insets. (E) Pearson’s correlation coefficient of GFP–LC3 vs. Lysotracker in control (10 cells), Tex2 KO-1 (10 cells), Tex2 KO-2 (10 cells), or Tex2 KO-1 treated with BafA1 (10 cells) in three independent assays. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. (F) Representative images of either control (top) or Tex2 KO-1 (bottom) cells expressing GFP-Lact2-C2 (green; PS sensor) with insets. (G) Pearson’s correlation coefficient of GFP-Lact2-C2 vs. Lamp1-mCh (18 control; 20 Tex2-KO cells). Two-tailed unpaired Student’s t test. Mean ± SD. (H) Representative images of either control (top) or Tex2 KO-1 (bottom) cells expressing GFP-P4M (PI4P sensor) with insets. (I) Pearson’s correlation coefficient of GFP-P4M vs. either Lamp1-mCh (13 control; 31 Tex2-KO cells) or B4GalT1-TM-BFP (Golgi marker; 13 control; 21 Tex2-KO cells). Two-tailed unpaired Student’s t test. Mean ± SD. (J) Linescan analysis of GFP-P4M at LE/lys/Golgi relative to the PM in control or Tex2 KO-1 HeLa cells, as shown in H. (K) Representative images of either control (top) or Tex2 KO-1 (bottom) cells expressing PI(4,5)P2 sensor PLCδ-PH-GFP with insets. (L) Linescan analysis of PLCδ-PH-GFP at LE/lys relative to the PM in control or Tex2 KO-1 HeLa cells, as shown in K. Two-tailed unpaired Student’s t test. Mean ± SD. Scale bar, 10 μm in the whole cell images and 2 μm in the insets in D, F, H, and K. Source data are available for this figure: SourceData FS5.

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