Loss of LAT1 sex-dependently delays recovery after caerulein-induced acute pancreatitis

Table 1 Antibodies and conditions applied in immunohistochemistry and immunofluorescence experiments

Antigen	Antigen retrieval	Primary antibody	Antibody dilution
Amylase	SDS 0.1%	LifeSpan BioSciences, Inc. (LS-B8239)	1:1000
Caspase 3	Sodium citrate pH 6	Cell Signaling (9664)	1:400
E-cadherin	Tris-EDTA pH 9	Invitrogen (13-1900)	1:200
LAT1	Tris-EDTA pH 9	Cosmo Bio (KE026)	1:100
pH3	Sodium citrate pH 6	Merk-Sigma (06-570)	1:1000
SMA	Sodium citrate pH 6	Abcam (AB5694)	1:500
SNAT5	Tris-EDTA pH 9	Pineda (599341)	1:2000

pH3: Phosphohistone 3; SMA: Smooth muscle actin.

Table 2 Antibodies and conditions applied in Western blotting experiments

Antigen	Total protein loaded (µg)	Primary antibody	Antibody dilution
Amylase	2.5	LifeSpan BioSciences, Inc. (LS-B8239)	1:10000
β-tubulin	Varying	Sigma (T8328)	1:5000
LAT1	20	Cosmobio (KE026)	1:500
LC3	100	MLB (PM036)	1:250
NA/K-ATPase	Varying	Santa Cruz (sc-48345)	1:10000
p62	50	MBL (PM045)	1:1000
peIF2α	25	Cell Signaling (3398)	1:1000
pS6K	50	Cell Signaling (9205S)	1:1000
SMA	25	Sigma (C6198)	1:1000
teIF2α	25	Cell Signaling (9722)	1:1000
tS6K	50	Cell Signaling (2708P)	1:1000
Vimentin	30	Invitrogen (MA5-11883)	1:1000

LC3: Microtubule-associated proteins 1A/1B light chain 3B; peIF2: Phosphorylated eukaryotic translation initiation factor 2; pS6K: Phosphorylated S6 kinase; SMA: Smooth muscle actin; teIF2α: Total eukaryotic translation initiation factor 2α; tS6K: Total S6 kinase.

Table 3 Primers and probes for mouse genes used in qPCR

Gene	Primers	Probe (Universal Probe Library Roche)	Accession No.
Amylase 2A4
S	TGATGACTGGGCTTTGTCAG	4	NM_001160150.1
AS	GCCATCGACCTTATCTCCAG
ccnb2
S	CAACCGTACCAAGTTCATCG	3	NM_007630.2
AS	GATGGCAGTCCAGTGTCTAGG
CD206
S	CCTTGAACCCATTTATCATTCC	49	NM_008625.2
AS	CTGTAGCAGTGGCCTGCAT
ck19
S	AGTCCCAGCTCAGCATGAA	97	NM_008471.3
AS	TAACGGGCCTCCGTCTCT
Clusterin
S	AAGTACTACCTTCGGGTCTCCA	6	NM_013492.3
AS	AGCTTCACCACCACCTCAGT
col4a1
S	GGCCCTTCATTAGCAGGTG	9	NM_009931.2
AS	TCTGAATGGTCTGACTGTGTACC
cpa1
S	CCCCTATGGCTACACATCAGA	63	NM_025350.4
AS	TCCCGTGTAGAGATGTCAAGG
ctsb
S	AAGCTGTGTGGCACTGTCCT	72	NM_007798.3
AS	GATCTATGTCCTCACCGAACG
ctsd
S	CCCTCCATTCATTGCAAGATAC	3	NM_009983.2
AS	TGCTGGACTTGTCACTGTTGT
ctsl
S	ACAGAAGACTGTATGGCACGAA	25	NM_009984.4
AS	GGATCATTCTCATGTTCTTCTCC
Elastase
S	ACGGAGCACCCTGTTACATC	22	NM_026419.2
AS	CTGCTGCAGCTTGTCTGG
F4/80
S	AGGAGGACTTCTCCAAGCCTA	67	NM_010130.4
AS	AGGCCTCTCAGACTTCTGCTT
icam-1
S	GCTACCATCACCGTGTATTCG	53	NM_010493.3
AS	AGGTCCTTGCCTACTTGCTG
IL-1β
S	AGTTGACGGACCCCAAAAG	38	NM_008361.4
AS	AGCTGGATGCTCTCATCAGG
IL-6
S	GCTACCAAACTGGATATAATCAGGA	6	NM_031168
AS	CCAGGTAGCTATGGTACTCCCGAA
IL-10
S	CAGAGCCACATGCTCCTAGA	41	NM_010548.2
AS	TGTCCAGCTGGTCCTTTGTT
lipase
S	GCCACGATGCTAATGCTGT	3	NM_026925.3
AS	ATCACTGAAGCAGCCGAGTT
mist1
S	GGCTAAAGCTACGTGTCCTTG	77	NM_010800.3
AS	GCTCCAGGCTGGTTTTCC
nox2
S	TGCCAACTTCCTCAGCTACA	20	NM_007807.5
AS	GTGCACAGCAAAGTGATTGG
p21
S	TTGCCAGCAGAATAAAAGGTG	9	NM_001111099.2
AS	TTTGCTCCTGTGCGGAAC
p27
S	GTTAGCGGAGCAGTGTCCA	62	NM_009875.4
AS	TCTGTTCTGTTGGCCCTTTT
p62
S	AACTGGAGCCCACGTCCT	22	NM_001290769.1
AS	CTCCCCCACATTCTTCAGG
prss1+3
S	CTGCTGGCACTCAGTGTCTC	20	NM_053243.2
AS	GCAGGTCTGGTTCACTGACA
prss2
S	CCAGAGTGGCCTCTGTACCT	20	NM_009430.2
AS	AGCAGGTCTGGGTTGTTCAC
ptf1a
S	GCTTGGCCATAGGCTACATT	73	NM_018809.2
AS	GGACTGTCCTCTCCGAGGT
rbpjl
S	TCAGAAATCCTATGGAAATGAGAA	20	NM_009036.1
AS	GGTCTTGCATTGGCTTCAC
sma/Acta2
S	TAACCCTTCAGCGTTCAGC	20	NM_007392.3
AS	ACATAGCTGGAGCAGCGTCT
sox9
S	CAGCAAGACTCTGGGCAAG	25	NM_011448.4
AS	ATCGGGGTGGTCTTTCTTGT
spink1
S	CACCCAGATCTTCGACAATG	83	NM_009258.5
AS	TAAACTCAGCAGGGCCAAAG
tnfα
S	CTGTAGCCCACGTCGTAGC	25	ENSMUST00000025263
AS	TTTGAGATCCATGCCGTTG
tgfβ
S	CTCAGAGCAAGAGAAAGCACTG	3	NM_001164074.1
AS	CGTTGATGAACCAGTTACAGACC
Vimentin
S	CGAGGAGAGCAGGATTTCTC	53	NM_011701.4
AS	GGAAGTGACTCCAGGTTAGTTTCTC

ccnb2: Cyclin b2; CD206/Mrc1: macrophage mannose receptor C type 1,ck19: Cytokeratin 19; col4a1: Collagen type IV alpha 1 chain; cpa1: Carpoxypeptidase 1; ctsb: Cathepsin b; ctsd: Cathepsin d; ctsl: Cathepsin l; icam-1: Intercellular adhesion molecule 1; IL-1: Interleukin 1; IL-6 and 10: Interleukin 6 and 10; mist1: Basic helix-loop-helix family member A15; nox2: NADPH oxidase 2; p21/cip/Cdkn1a: Cyclin Dependent Kinase Inhibitor 1A; p27/kip/Cdkn1b: cyclin-dependent kinase inhibitor 1B, prss1+3: Serine protease 1+3; prss2: Serine protease 2; ptf1a: Pancreas associated transcription factor 1a; rbpjl: Recombining binding protein suppressor of hairless-like protein; sma/Acta2: α smooth muscle actin/alpha-actin-2; sox9: SRY-box transcription factor 9; spink1: Serine peptidase inhibitor kazal type 1; tnfα: Tumor necrosis factor α; tgfβ: Transforming growth factor β.