Journal: Developmental cell

Article Title: EGFR/Pak Signaling Selectively Regulates Glutamine Deprivation-Induced Macropinocytosis

doi: 10.1016/j.devcel.2019.05.043

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Compound, Sakura Finetek VWR Cat#25608-930 Dako Fluorescence Mounting Medium Agilent Tech Cat#S302380-2 PureLink RNA Mini Kit Thermo Fisher Scientific Cat#12183025 High Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat#4368814 SYBR Premix Ex Taq II (Tli RNaseH Plus) TaKaRa Cat#RR820L DC Protein Assay Kit Bio-Rad Cat#5000112 NewBlot IR Stripping Buffer, 5X LI-COR Cat#928-40028 Lipofectamine RNAiMAX Transfection Reagent Thermo Fisher Scientific Cat#13778100 VECTASTAIN Elite ABC HRP Kit Vector Laboratories Cat#PK-6100 DAB HRP Substrate Kit Vector Laboratories Cat#PK-4100 Experimental Models: Cell Lines AsPC-1 ATCC ATCC CRL-1682 HPAF-II ATCC ATCC CRL-1997 SW1990 ATCC ATCC CRL-2172 MIA PaCa-2 (MP-2) ATCC ATCC CRL-1420 Panc-1 ATCC ATCC CRL-1469 CFPAC-1 ATCC ATCC CRL-1918 Experimental Models: Organisms/Strains Mouse: NCr nude, sp/sp, CrTac:NCr-Foxn1nu Taconic Cat#NCRNU-F Mouse: Foxn1 nu /Foxn1 nu Jackson Lab Cat#002019 Oligonucleotides AREG Forward: 5′-CGGGAGCCGACTATGACTAC-3′ Reverse: 5′-TCACTTTCCGTCTTGTTTTGG-3′ This paper N/A BTC Forward: 5′-AGAGCGGGGTTGATGGAC-3′ Reverse: 5′-ATCTGCCACCACACAGTGAA-3′ This paper N/A EGF Forward: 5′-GTACTCTCGCAGGAAATGGG-3′ Reverse: 5′-TCCACCACCAATTGCTCATA-3′ This paper N/A EPGN Forward: 5′-ACTACAGGAAATGGCTTTGGG-3′ Reverse: 5′-GGCTATGGGTCCTTCTATGTTG-3′ This paper N/A EREG Forward: 5′-AGGAGGATGGAGATGCTCTG-3′ Reverse: 5′-CACAGTTGTACTGAGGACTGCC-3′ This paper N/A HBEGF Forward: 5′-GGACCCATGTCTTCGGAAAT-3′ Reverse: 5′-CCCATGACACCTCTCTCCAT-3′ This paper N/A TGFA Forward: 5′-CCTGTTCGCTCTGGGTATTG-3′ Reverse: 5′-ACTGAGTGTGGGAATCTGGG-3′ This paper N/A 18S rRNA Forward: 5′-GTAACCCGTTGAACCCCATT-3′ Reverse: 5′-CCATCCAATCGGTAGTAGCG-3′ This paper N/A Silencer Select Negative Control No. 1 siRNA Thermo Fisher Scientific Cat#4390843 Silencer Select EGFR siRNA #1 Thermo Fisher Scientific ID#s563 Silencer Select EGFR siRNA #2 Thermo Fisher Scientific ID#s565 Silencer Select KRAS siRNA #1 Thermo Fisher Scientific ID#s7939 Silencer Select KRAS siRNA #2 Thermo Fisher Scientific ID#s7940 Software and Algorithms ImageJ software NIH https://imagej.nih.gov/ij/ GraphPad Prism 8 software GraphPad https://www.graphpad.com/ Image Studio Lite software LI-COR https://www.licor.com/bio/ Open in a separate window KEY RESOURCES TABLE siRNA Transfection Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher Scientific) was used with a final siRNA concentration of 25 nM according to the manufacturer’s protocol.

Techniques: Recombinant, Fluorescence, DC Protein Assay, Stripping Membranes, Transfection, Plasmid Preparation, Negative Control, Software