Figure S3.
Tamoxifen-inducible knockdown of mDia1 and ARPC2. (A) Mouse expressing Cre recombinase under the acinar-specific Mist1 TMX-inducible promoter and the membrane reporter mTom/mGFP (CtrlMist1-Cre, mT/mG). Mice were treated with 75 mg/kg TMX for 4 wk and salivary glands exposed and imaged by confocal microscopy. Acinar cells positive for GFP express Cre recombinase. Bar, 10 µm. (B) Mice in which exon 1 of the gene encoding mDia1 (DIAPH1) is flanked by LoxP sites were crossed with the acinar-specific Mist1 TMX-inducible promoter/membrane reporter mTom/mGFP mice (mDia1fl/fl Mist1-Cre, mT/mG). After 4 wk of TMX treatment, salivary glands were exposed, processed for indirect immunofluorescence, and labeled with antibodies directed against mDia1. Bar, 10 µm. Insets show the variability of mdia1 levels on different granules. Bar, 1 µm. The graph shows the levels of mDia1 in mGFP and mTom cells both in mDia1fl/fl Mist1-Cre, mT/mG mice and CtrlMist1-Cre, mT/mG mice used as additional control. Levels were expressed as mean fluorescence intensity per animal, as described in Materials and methods (70 cells per group, 3 animals per group). (C) Mice in which exon 8 of the gene encoding Arpc2 (ARPC2) is flanked by LoxP sites were crossed with the acinar-specific Mist1 TMX-inducible promoter/membrane reporter mTom/mGFP mice (ARPC2fl/fl Mist1-Cre, mT/mG mG). After 4 wk of TMX treatment, mice were injected with 0.01 mg/kg ISOP, and salivary glands were excised, processed for indirect immunofluorescence, and labeled with antibodies directed against ARPC2. Cells that express Cre recombinase (green and highlighted by dashed lines) have lower levels of ARPC2 than those that do not (magenta). Bar, 10 µm. Insets show that in cells that do not express Cre (green), ARPC2 is not present on the fused granules. Bar, 1 µm. The graph shows the levels of ARPC2 in mGFP and mTom cells both in ARPC2fl/fl Mist1-Cre, mT/mG and CtrlMist1-Cre, mT/mG mice. Levels were expressed as mean fluorescence intensity per animal, as described in Materials and methods (70 cells per group, 3 animals per group).

Tamoxifen-inducible knockdown of mDia1 and ARPC2. (A) Mouse expressing Cre recombinase under the acinar-specific Mist1 TMX-inducible promoter and the membrane reporter mTom/mGFP (CtrlMist1-Cre, mT/mG). Mice were treated with 75 mg/kg TMX for 4 wk and salivary glands exposed and imaged by confocal microscopy. Acinar cells positive for GFP express Cre recombinase. Bar, 10 µm. (B) Mice in which exon 1 of the gene encoding mDia1 (DIAPH1) is flanked by LoxP sites were crossed with the acinar-specific Mist1 TMX-inducible promoter/membrane reporter mTom/mGFP mice (mDia1fl/fl Mist1-Cre, mT/mG). After 4 wk of TMX treatment, salivary glands were exposed, processed for indirect immunofluorescence, and labeled with antibodies directed against mDia1. Bar, 10 µm. Insets show the variability of mdia1 levels on different granules. Bar, 1 µm. The graph shows the levels of mDia1 in mGFP and mTom cells both in mDia1fl/fl Mist1-Cre, mT/mG mice and CtrlMist1-Cre, mT/mG mice used as additional control. Levels were expressed as mean fluorescence intensity per animal, as described in Materials and methods (70 cells per group, 3 animals per group). (C) Mice in which exon 8 of the gene encoding Arpc2 (ARPC2) is flanked by LoxP sites were crossed with the acinar-specific Mist1 TMX-inducible promoter/membrane reporter mTom/mGFP mice (ARPC2fl/fl Mist1-Cre, mT/mG mG). After 4 wk of TMX treatment, mice were injected with 0.01 mg/kg ISOP, and salivary glands were excised, processed for indirect immunofluorescence, and labeled with antibodies directed against ARPC2. Cells that express Cre recombinase (green and highlighted by dashed lines) have lower levels of ARPC2 than those that do not (magenta). Bar, 10 µm. Insets show that in cells that do not express Cre (green), ARPC2 is not present on the fused granules. Bar, 1 µm. The graph shows the levels of ARPC2 in mGFP and mTom cells both in ARPC2fl/fl Mist1-Cre, mT/mG and CtrlMist1-Cre, mT/mG mice. Levels were expressed as mean fluorescence intensity per animal, as described in Materials and methods (70 cells per group, 3 animals per group).

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