Table 1.
Antibodies, siRNAs, primers, and DNA constructs used in this study
Reagent or resourceSourceIdentifier
Antibodies   
Rabbit anti-MASTL (1:1,000 for Western blot [WB]) Sigma-Aldrich Cat# HPA027175; RRID:AB_1853591 
Rabbit anti-MASTL (1:1,000 for WB) Abcam Cat# ab86387; RRID:AB_1925198 
Mouse anti-α-tubulin (1:1,000 for WB, 1:100 for immunofluorescence [IF]) Hybridoma bank 12g10anti-alpha-tubulin 
Mouse anti-β1-integrin CD29 (1:1,000 for WB) BD Biosciences Cat# 610468; RRID:AB_397840 
Mouse anti-GAPDH (1:20,000 for WB) HyTest Cat# 5G4MAB6C5 
Mouse anti-paxillin (1:100 for IF, 1:1,000 for WB) BD Biosciences Cat# 612405, RRID:AB_647289 
Rabbit anti-phospho-MLC 2 (Thr18/Ser19; 1:100 for IF) Cell Signaling Technology Cat# 3674, RRID:AB_2147464 
Rabbit anti-MLC (phospho S20; 1:500 for WB) Abcam Cat# ab2480, RRID:AB_303094 
Rabbit anti-MLC 2 (1:500 for WB) Cell Signaling Technology Cat# 3672, RRID:AB_10692513 
Rabbit anti-GEF-H1 (1:1,000 for WB) Cell Signaling Technology Cat# 4076, RRID:AB_2060032 
Rabbit anti-phospho-GEF-H1 (Ser886; 1:1,000 for WB) Cell Signaling Technology Cat# 14143, RRID:AB_2798402 
Rabbit anti-tropomyosin 4.2 (Delta Tm 9d; 1:1,000 for WB, 1:100 for IF) A kind gift from Prof. Peter Gunning (UNSW Sydney, Sydney, Australia) (Schevzov et al., 2011) 
Mouse anti-nonmuscle myosin IIB (1:1,000 for WB) Abcam Cat# ab684, RRID:AB_305661 
Mouse anti-vinculin (1:100 for IF, 1:1,000 for WB) Sigma-Aldrich Cat# V9131, RRID:AB_477629 
Mouse anti-β-actin (clone AC-15; 1:1,000 for WB) Sigma-Aldrich Cat# A1978, RRID:AB_476692 
Recombinant anti-β-catenin antibody (E247; 1:100 for IF) Abcam Cat# ab32572, RRID:AB_725966 
Rabbit anti-E-cadherin (24E10; 1:100 for IF) Cell Signaling Technology Cat# 3195, RRID:AB_2291471 
Rabbit anti-GEF-H1 (IF 1:100) Abcam Cat# ab155785, RRID:AB_2818944 
Rabbit anti-phospho-paxillin (Tyr118; 1:1,000 for WB) Cell Signaling Technology Cat# 2541, RRID:AB_2174466 
Mouse anti-MRTF-A (G-8; 1:200 for IF) Santa Cruz Biotechnology Cat# sc-390324 
Donkey anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody, Alexa Fluor 568 (1:400 for IF) Thermo Fisher Scientific Cat# A10037, RRID:AB_2534013 
Donkey anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody, Alexa Fluor 488 (1:400 for IF) Invitrogen Cat# A-21202 
IRDye 680RD donkey anti-mouse IgG (1:5,000 for WB) LI-COR Biosciences Cat# 926-68072 
Donkey anti-rabbit IgG (H+L) highly cross-adsorbed secondary antibody, Alexa Fluor 488 (1:400 for IF) Invitrogen Cat# A-21206 
IRDye 800CW donkey anti-rabbit IgG (1:5,000 for WB) LI-COR Biosciences Cat# 926-32213 
siRNAs and primers 
AllStars negative control siRNA Qiagen 1027281 
Hs_MASTL_6 FlexiTube siRNA (target sequence 5′-ACG​CCT​TAT​TCT​AGC​AAA​TTA-3′) Qiagen SI02653014 
Hs_MASTL_7 FlexiTube siRNA (target sequence 5′-CAG​GAC​AAG​TGT​TAT​CGC​TTA-3′) Qiagen SI02653182 
Hs_MASTL_11 FlexiTube siRNA (target sequence 5′-CAG​CCC​TTA​GAT​TCA​GAT​AGA-3′) Qiagen SI04441066 
Hs_MKL1_7 FlexiTube siRNA (target sequence 5′-ATC​ACG​TGT​GAT​TGA​CAT​GTA-3′) Qiagen SI04172028 
MRTF-A (sense strand 5′-CCA​UCA​UUG​UGG​GCC​AGG​U-3′) Sigma-Aldrich PDSIRNA5D 
Hs_SRF_5 FlexiTube siRNA (target sequence 5′-CAA​GAT​GGA​GTT​CAT​CGA​CAA-3′) Qiagen SI02757622 
 RT-PCR primer for ARHGEF2 Universal Probe Library (Roche) 
Forward: 5′-TAC​CTG​CGG​CGA​ATT​AAG​AT-3′ 
Reverse: 5′-AAA​CAG​CCC​GAC​CTT​CTC​TC-3′ 
Probe: 22 
 RT-PCR primer for GAPDH Universal Probe Library (Roche) 
Forward: 5′-GCC​CAA​TAC​GAC​CAA​ATC​C-3′ 
Reverse: 5′-AGC​CAC​ATC​GCT​CAG​ACA-3′ 
Probe: 60 
 RT-PCR primer for MYH10 Universal Probe Library (Roche) 
Forward: 5′-GGA​GGA​CAC​GCT​GGA​CAC-3′ 
Reverse: 5′-TCT​GCC​ACT​TCT​TGT​TCA​CG-3′ 
Probe: 3 
 RT-PCR primer for MASTL Universal Probe Library (Roche) 
Forward: 5′-GTC​TGC​AAA​CAA​TGT​CTA​CTT​GGT​A-3′ 
Reverse: 5′-CTA​GAG​CCA​GTG​CTA​CTT​CAG​AAA-3′ 
Probe: 76 
 RT-PCR primer for TPM4 Universal Probe Library (Roche) 
Forward: 5′-CGG​TGA​AAC​GCA​AGA​TCC-3′ 
Reverse: 5′-GGC​CAC​ATC​ACC​TTC​AGC-3′ 
Probe: 24 
 qPCR primer for TBP N/A N/A 
Forward: 5′-TGC​ACA​GGA​GCC​AAG​AGT​GAA-3′ 
Reverse: 5′-CAC​ATC​ACA​GCT​CCC​CAC​CA-3′ 
 qPCR primer for SRF N/A N/A 
Forward: 5′-CAG​ATC​GGT​ATG​GTG​GTC​GG-3′ 
Reverse: 5′-GTC​AGC​GTG​GAC​AGC​TCA​TA-3′ 
 qPCR primer for FOS N/A N/A 
Forward: 5′-CTC​TCT​TAC​TAC​CAC​TCA​CCC​GC-3′ 
Reverse: 5′-GGT​CCG​TGC​AGA​AGT​CCT​GCG-3′ 
 qPCR primer for VCL N/A N/A 
Forward: 5′-ACC​AAG​GCA​TAG​AGG​AAG​CTT​TA-3′ 
Reverse: 5′-GCT​CTC​TTC​ATG​GCT​TCA​GTG-3′ 
DNA constructs 
pcDNA EGFP-C2 This paper N/A 
pcDNA EGFP MASTL WT (siRNA resistant) This paper N/A 
pcDNA EGFP MASTL G44S (siRNA resistant) This paper N/A 
pcDNA EGFP MASTL E167D (siRNA resistant) This paper N/A 
pIND20-MRTF-A-GFP Hinojosa et al., 2017  N/A 
mEmerald-paxillin-22 A kind gift from Prof. Robert Grosse, Institute of Experimental and Clinical Pharmacology and Toxicology, University of Freiburg and CIBSS – Center for Integrative Biological Signalling Studies, Freiburg, Germany (plasmid 54219; Addgene; http://n2t.net/addgene:54219; RRID:Addgene_54219) (Paszek et al., 2012) 
pcDNA3.1 EGFP ARHGEF2 A kind gift from Prof. Michael Sixt (The Institute of Science and Technology Austria, Klosterneuburg, Austria) Kopf et al., 2019 Preprint  
Reagent or resourceSourceIdentifier
Antibodies   
Rabbit anti-MASTL (1:1,000 for Western blot [WB]) Sigma-Aldrich Cat# HPA027175; RRID:AB_1853591 
Rabbit anti-MASTL (1:1,000 for WB) Abcam Cat# ab86387; RRID:AB_1925198 
Mouse anti-α-tubulin (1:1,000 for WB, 1:100 for immunofluorescence [IF]) Hybridoma bank 12g10anti-alpha-tubulin 
Mouse anti-β1-integrin CD29 (1:1,000 for WB) BD Biosciences Cat# 610468; RRID:AB_397840 
Mouse anti-GAPDH (1:20,000 for WB) HyTest Cat# 5G4MAB6C5 
Mouse anti-paxillin (1:100 for IF, 1:1,000 for WB) BD Biosciences Cat# 612405, RRID:AB_647289 
Rabbit anti-phospho-MLC 2 (Thr18/Ser19; 1:100 for IF) Cell Signaling Technology Cat# 3674, RRID:AB_2147464 
Rabbit anti-MLC (phospho S20; 1:500 for WB) Abcam Cat# ab2480, RRID:AB_303094 
Rabbit anti-MLC 2 (1:500 for WB) Cell Signaling Technology Cat# 3672, RRID:AB_10692513 
Rabbit anti-GEF-H1 (1:1,000 for WB) Cell Signaling Technology Cat# 4076, RRID:AB_2060032 
Rabbit anti-phospho-GEF-H1 (Ser886; 1:1,000 for WB) Cell Signaling Technology Cat# 14143, RRID:AB_2798402 
Rabbit anti-tropomyosin 4.2 (Delta Tm 9d; 1:1,000 for WB, 1:100 for IF) A kind gift from Prof. Peter Gunning (UNSW Sydney, Sydney, Australia) (Schevzov et al., 2011) 
Mouse anti-nonmuscle myosin IIB (1:1,000 for WB) Abcam Cat# ab684, RRID:AB_305661 
Mouse anti-vinculin (1:100 for IF, 1:1,000 for WB) Sigma-Aldrich Cat# V9131, RRID:AB_477629 
Mouse anti-β-actin (clone AC-15; 1:1,000 for WB) Sigma-Aldrich Cat# A1978, RRID:AB_476692 
Recombinant anti-β-catenin antibody (E247; 1:100 for IF) Abcam Cat# ab32572, RRID:AB_725966 
Rabbit anti-E-cadherin (24E10; 1:100 for IF) Cell Signaling Technology Cat# 3195, RRID:AB_2291471 
Rabbit anti-GEF-H1 (IF 1:100) Abcam Cat# ab155785, RRID:AB_2818944 
Rabbit anti-phospho-paxillin (Tyr118; 1:1,000 for WB) Cell Signaling Technology Cat# 2541, RRID:AB_2174466 
Mouse anti-MRTF-A (G-8; 1:200 for IF) Santa Cruz Biotechnology Cat# sc-390324 
Donkey anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody, Alexa Fluor 568 (1:400 for IF) Thermo Fisher Scientific Cat# A10037, RRID:AB_2534013 
Donkey anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody, Alexa Fluor 488 (1:400 for IF) Invitrogen Cat# A-21202 
IRDye 680RD donkey anti-mouse IgG (1:5,000 for WB) LI-COR Biosciences Cat# 926-68072 
Donkey anti-rabbit IgG (H+L) highly cross-adsorbed secondary antibody, Alexa Fluor 488 (1:400 for IF) Invitrogen Cat# A-21206 
IRDye 800CW donkey anti-rabbit IgG (1:5,000 for WB) LI-COR Biosciences Cat# 926-32213 
siRNAs and primers 
AllStars negative control siRNA Qiagen 1027281 
Hs_MASTL_6 FlexiTube siRNA (target sequence 5′-ACG​CCT​TAT​TCT​AGC​AAA​TTA-3′) Qiagen SI02653014 
Hs_MASTL_7 FlexiTube siRNA (target sequence 5′-CAG​GAC​AAG​TGT​TAT​CGC​TTA-3′) Qiagen SI02653182 
Hs_MASTL_11 FlexiTube siRNA (target sequence 5′-CAG​CCC​TTA​GAT​TCA​GAT​AGA-3′) Qiagen SI04441066 
Hs_MKL1_7 FlexiTube siRNA (target sequence 5′-ATC​ACG​TGT​GAT​TGA​CAT​GTA-3′) Qiagen SI04172028 
MRTF-A (sense strand 5′-CCA​UCA​UUG​UGG​GCC​AGG​U-3′) Sigma-Aldrich PDSIRNA5D 
Hs_SRF_5 FlexiTube siRNA (target sequence 5′-CAA​GAT​GGA​GTT​CAT​CGA​CAA-3′) Qiagen SI02757622 
 RT-PCR primer for ARHGEF2 Universal Probe Library (Roche) 
Forward: 5′-TAC​CTG​CGG​CGA​ATT​AAG​AT-3′ 
Reverse: 5′-AAA​CAG​CCC​GAC​CTT​CTC​TC-3′ 
Probe: 22 
 RT-PCR primer for GAPDH Universal Probe Library (Roche) 
Forward: 5′-GCC​CAA​TAC​GAC​CAA​ATC​C-3′ 
Reverse: 5′-AGC​CAC​ATC​GCT​CAG​ACA-3′ 
Probe: 60 
 RT-PCR primer for MYH10 Universal Probe Library (Roche) 
Forward: 5′-GGA​GGA​CAC​GCT​GGA​CAC-3′ 
Reverse: 5′-TCT​GCC​ACT​TCT​TGT​TCA​CG-3′ 
Probe: 3 
 RT-PCR primer for MASTL Universal Probe Library (Roche) 
Forward: 5′-GTC​TGC​AAA​CAA​TGT​CTA​CTT​GGT​A-3′ 
Reverse: 5′-CTA​GAG​CCA​GTG​CTA​CTT​CAG​AAA-3′ 
Probe: 76 
 RT-PCR primer for TPM4 Universal Probe Library (Roche) 
Forward: 5′-CGG​TGA​AAC​GCA​AGA​TCC-3′ 
Reverse: 5′-GGC​CAC​ATC​ACC​TTC​AGC-3′ 
Probe: 24 
 qPCR primer for TBP N/A N/A 
Forward: 5′-TGC​ACA​GGA​GCC​AAG​AGT​GAA-3′ 
Reverse: 5′-CAC​ATC​ACA​GCT​CCC​CAC​CA-3′ 
 qPCR primer for SRF N/A N/A 
Forward: 5′-CAG​ATC​GGT​ATG​GTG​GTC​GG-3′ 
Reverse: 5′-GTC​AGC​GTG​GAC​AGC​TCA​TA-3′ 
 qPCR primer for FOS N/A N/A 
Forward: 5′-CTC​TCT​TAC​TAC​CAC​TCA​CCC​GC-3′ 
Reverse: 5′-GGT​CCG​TGC​AGA​AGT​CCT​GCG-3′ 
 qPCR primer for VCL N/A N/A 
Forward: 5′-ACC​AAG​GCA​TAG​AGG​AAG​CTT​TA-3′ 
Reverse: 5′-GCT​CTC​TTC​ATG​GCT​TCA​GTG-3′ 
DNA constructs 
pcDNA EGFP-C2 This paper N/A 
pcDNA EGFP MASTL WT (siRNA resistant) This paper N/A 
pcDNA EGFP MASTL G44S (siRNA resistant) This paper N/A 
pcDNA EGFP MASTL E167D (siRNA resistant) This paper N/A 
pIND20-MRTF-A-GFP Hinojosa et al., 2017  N/A 
mEmerald-paxillin-22 A kind gift from Prof. Robert Grosse, Institute of Experimental and Clinical Pharmacology and Toxicology, University of Freiburg and CIBSS – Center for Integrative Biological Signalling Studies, Freiburg, Germany (plasmid 54219; Addgene; http://n2t.net/addgene:54219; RRID:Addgene_54219) (Paszek et al., 2012) 
pcDNA3.1 EGFP ARHGEF2 A kind gift from Prof. Michael Sixt (The Institute of Science and Technology Austria, Klosterneuburg, Austria) Kopf et al., 2019 Preprint  
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