Supplemental material (PDF)
SUPPLEMENTAL MATERIAL Matin et al., http://www.jem.org/cgi/content/full/jem.20121439/DC1
Table S1.
Statistical analysis of demographic data for p63-expressing primary mTMA samples
The Jour nal of Exper imental Medicine
Demographic data Mean age in years (SD) Gender (male/female) Site of melanoma Trunk Limbs Acral Mucosal
p63 negative (n = 24)
p63 positive (n = 16)
54 (16.5) 12:12
56.3 (14.4) 10:6
6 4 11 3
9 3 2 2
P-value 0.64 (Student’s t test) 0.33 (2)
0.044 (2)
[ID]TBLS1/
[ID]TBLS2/
[ID]TBLS3/
[ID]TBLS4/
[ID]TBLS5/
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Table S2.
Statistical analysis of clinicopathological data for primary melanoma archival tissue samples
Clinicopathological factors
p63 negative (n = 34)
p63 positive (n = 46)
Total (n = 80)
P-value
Age at diagnosis (yr) Gender Male Female Site of melanoma Acral Extremities Head/Neck Trunk Breslow thickness T1 T2 T3 T4 Clark level I II III IV V Missing Histological classification ALM NMM SMM Growth phase RGP VGP Missing Ulceration status No Yes Missing Mitotic rate Stage 1 Stage 2 Stage 3 Stage 4 Missing Regression No Yes Missing Microsatellites No Yes Missing
58.0 (46.3, 74.7)
64.1 (40.1, 72.4)
61.0 (44.1, 73.3)
0.81
14 (41.2) 20 (58.8)
21 (45.7) 25 (54.3)
35 (43.8) 45 (56.3)
0.69
4 (11.8) 12 (35.3) 4 (11.8) 14 (41.2)
7 (15.2) 13 (28.3) 9 (19.6) 17 (37.0)
11 (13.8) 25 (31.3) 13 (16.3) 31 (38.8)
0.52 0.77 0.61
14 (41.2) 5 (14.7) 10 (29.4) 5 (14.7)
11 (23.9) 7 (15.2) 10 (21.7) 18 (39.1)
25 (31.3) 12 (15.0) 20 (25.0) 23 (28.8)
0.03
0 9 (27.3) 8 (24.2) 14 (42.4) 2 (6.1) 1
0 7 (15.6) 12 (26.7) 20 (44.4) 6 (13.3) 1
0 16 (20.5) 20 (25.6) 34 (43.6) 8 (10.3) 2
4 (11.8) 12 (35.3) 18 (52.9)
5 (10.9) 22 (47.8) 19 (41.3)
9 (11.3) 34 (42.5) 37 (46.3)
0.61 0.82
8 (25.0) 24 (75.0) 2
8 (17.8) 37 (82.2) 1
16 (20.8) 61 (79.2) 3
0.44
23 (69.7) 10 (30.3) 1
25 (54.4) 21 (45.7) 0
48 (60.8) 31 (39.2) 1
0.17
9 (32.1) 7 (25.0) 8 (25.6) 4 (14.3) 6
12 (34.3) 8 (22.9) 14 (40.0) 1 (2.9) 11
21 (33.3) 15 (23.8) 22 (34.9) 5 (7.9) 17
0.59
29 (87.9) 4 (12.1) 1
35 (77.8) 10 (22.2) 1
64 (82.1) 14 (18.0) 2
0.25
28 (84.9) 5 (15.2) 1
41 (91.1) 4 (8.9) 1
69 (88.5) 9 (11.5) 2
0.39
0.19
ALM, acral lentiginous melanoma; NMM, nodular melanoma; SSM, superficial spreading melanoma; VGP, vertical growth phase. Numbers are number (percentage) or median (interquartile range) as appropriate. P-values are odds comparing p63 negative with p63 positive. “-” indicates the parameter against which the regression analysis was taken; therefore, there is no p-value for this parameter.
S2
Oncogenic p63 is a p53 repressor in melanoma | Matin et al.
Table S3.
Statistical analysis of clinicopathological data for metastatic melanoma archival tissue samples
Clinicopathological factors Age at diagnosis (yr) Gender Male Female Site of metastasis Lymph Other sitesa
p63 negative (n = 28)
p63 positive (n = 47)
Total (n = 75)
P-value
62.0 (14.9)
60.2 (16.1)
60.9 (15.6)
0.69
19 9
23 24
42 33
0.19
9 19
24 23
33 42
0.15
Numbers are number (percentage) or median (interquartile range) as appropriate. P-values are odds comparing p63 negative with p63 positive. aSkin, lung, parotid, and brain.
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Table S4. Mutational status of p53, BRAF, and NRAS in melanoma cell lines
Table S5. Culture media and supplements for the melanocytes and established melanoma cell lines
Cell line
Cell line
TP53 status
RGP cells WM35 WT WM1575 WT WM1552C R248Q/+ SBCl2 WT VGP cells WM793 WT WM115 WT WM278 WT VMM39 WT LM3 WT LM4 Y236H/++ LM5 WT Mel 224 WT Mel 505 WT Metastatic melanoma cells LM32 WT LM34 WT LM37 WT Mel 501 WT SK-MEL 5 WT DX3 P278R/+ DX3LT5.1 P278R/+ WM1158 WT WM9 WT WM852 S241P WM983B mt A375M WT C8161 WT SK-MEL 31 WT MALME-3M WT SK-MEL 24 WT HBL WT WM239A WT MM-AN Null UISO-mel-6 Null VGP, vertical growth phase.
BRAF status
NRAS status
V600E V600E V600E WT
WT WT WT Q61K
V600E V600D V600E WT WT V600E V600E WT WT
WT WT WT WT Q61R WT WT Q61R WT
WT V600E WT V600E WT WT WT V600E V600E WT V600E V600E WT WT V600E V600E WT V600D V600E WT
WT WT WT WT WT Q61R Q61R WT WT Q61R WT WT WT WT WT WT WT WT WT Q61R
Culture media and supplements
Human primary melanocytes NHEM, HEMa 254 medium + HMGS-2 (Invitrogen) RGP cells WM35 RPMI + 10% FCS + 1% Gln WM1575 Tu 2% + 10% FCS WM1552C Tu 2% + 10% FCS SBCl2 RPMI + 10% FCS + 1% Gln VGP cells WM793 RPMI + 10% FCS + 1% Gln WM115 RPMI + 10% FCS + 1% Gln WM278 RPMI + 10% FCS + 1% Gln VMM39 RPMI + 10% FCS + 1% Gln LM3 RPMI + 10% FCS + 1% Gln LM4 RPMI + 10% FCS + 1% Gln LM5 RPMI + 10% FCS + 1% Gln Mel 224 RPMI + 10% FCS + 0.5% NEAA + 1% Gln Mel 505 RPMI + 10% FCS + 0.5% NEAA + 1% Gln Metastatic melanoma cells LM32 RPMI + 10% FCS + 1% Gln LM34 RPMI + 10% FCS + 1% Gln LM37 RPMI + 10% FCS + 1% Gln Mel 501 F10 + 10% FCS + 1% Gln SK- MEL 5 MEM + 10% FCS + 1% Gln DX3 RPMI + 10% FCS + 1% Gln DX3-LT5.1 RPMI + 10% FCS + 1% Gln WM1158 RPMI + 10% FCS + 1% Gln WM9 RPMI + 10% FCS + 1% Gln WM852 RPMI + 10% FCS + 1% Gln WM983B RPMI + 10% FCS + 1% Gln A375M RPMI + 10% FCS + 1% Gln C8161 RPMI + 10% FCS+ 20 mM Hepes + 0.1% GS SK-MEL 31 MEM + 10% FCS + 1% Gln MALME-3M RPMI+ 10% FCS + 1% Gln SK-MEL 24 MEM + 10% FCS + 1% Gln HBL MEM + 10% FCS + 1 mM SP + 1% Gln WM239A RPMI + 10% FCS + 1% Gln MM-AN DMEM + 10% FCS + 1% Gln UISO-mel-6 MEM + 15% FCS+ 0.5% NEAA + 1% Gln + 0.1% GS HMGS-2, human melanocytes growth supplement-2; Gln, glutamine; GS, gentamicin sulphate; NEAA, nonessential amino acids; SP, sodium pyruvate; VGP, vertical growth phase.
S4
Oncogenic p63 is a p53 repressor in melanoma | Matin et al.
Table S1.
Statistical analysis of demographic data for p63-expressing primary mTMA samples
The Jour nal of Exper imental Medicine
Demographic data Mean age in years (SD) Gender (male/female) Site of melanoma Trunk Limbs Acral Mucosal
p63 negative (n = 24)
p63 positive (n = 16)
54 (16.5) 12:12
56.3 (14.4) 10:6
6 4 11 3
9 3 2 2
P-value 0.64 (Student’s t test) 0.33 (2)
0.044 (2)
[ID]TBLS1/
[ID]TBLS2/
[ID]TBLS3/
[ID]TBLS4/
[ID]TBLS5/
JEM
S1
Table S2.
Statistical analysis of clinicopathological data for primary melanoma archival tissue samples
Clinicopathological factors
p63 negative (n = 34)
p63 positive (n = 46)
Total (n = 80)
P-value
Age at diagnosis (yr) Gender Male Female Site of melanoma Acral Extremities Head/Neck Trunk Breslow thickness T1 T2 T3 T4 Clark level I II III IV V Missing Histological classification ALM NMM SMM Growth phase RGP VGP Missing Ulceration status No Yes Missing Mitotic rate Stage 1 Stage 2 Stage 3 Stage 4 Missing Regression No Yes Missing Microsatellites No Yes Missing
58.0 (46.3, 74.7)
64.1 (40.1, 72.4)
61.0 (44.1, 73.3)
0.81
14 (41.2) 20 (58.8)
21 (45.7) 25 (54.3)
35 (43.8) 45 (56.3)
0.69
4 (11.8) 12 (35.3) 4 (11.8) 14 (41.2)
7 (15.2) 13 (28.3) 9 (19.6) 17 (37.0)
11 (13.8) 25 (31.3) 13 (16.3) 31 (38.8)
0.52 0.77 0.61
14 (41.2) 5 (14.7) 10 (29.4) 5 (14.7)
11 (23.9) 7 (15.2) 10 (21.7) 18 (39.1)
25 (31.3) 12 (15.0) 20 (25.0) 23 (28.8)
0.03
0 9 (27.3) 8 (24.2) 14 (42.4) 2 (6.1) 1
0 7 (15.6) 12 (26.7) 20 (44.4) 6 (13.3) 1
0 16 (20.5) 20 (25.6) 34 (43.6) 8 (10.3) 2
4 (11.8) 12 (35.3) 18 (52.9)
5 (10.9) 22 (47.8) 19 (41.3)
9 (11.3) 34 (42.5) 37 (46.3)
0.61 0.82
8 (25.0) 24 (75.0) 2
8 (17.8) 37 (82.2) 1
16 (20.8) 61 (79.2) 3
0.44
23 (69.7) 10 (30.3) 1
25 (54.4) 21 (45.7) 0
48 (60.8) 31 (39.2) 1
0.17
9 (32.1) 7 (25.0) 8 (25.6) 4 (14.3) 6
12 (34.3) 8 (22.9) 14 (40.0) 1 (2.9) 11
21 (33.3) 15 (23.8) 22 (34.9) 5 (7.9) 17
0.59
29 (87.9) 4 (12.1) 1
35 (77.8) 10 (22.2) 1
64 (82.1) 14 (18.0) 2
0.25
28 (84.9) 5 (15.2) 1
41 (91.1) 4 (8.9) 1
69 (88.5) 9 (11.5) 2
0.39
0.19
ALM, acral lentiginous melanoma; NMM, nodular melanoma; SSM, superficial spreading melanoma; VGP, vertical growth phase. Numbers are number (percentage) or median (interquartile range) as appropriate. P-values are odds comparing p63 negative with p63 positive. “-” indicates the parameter against which the regression analysis was taken; therefore, there is no p-value for this parameter.
S2
Oncogenic p63 is a p53 repressor in melanoma | Matin et al.
Table S3.
Statistical analysis of clinicopathological data for metastatic melanoma archival tissue samples
Clinicopathological factors Age at diagnosis (yr) Gender Male Female Site of metastasis Lymph Other sitesa
p63 negative (n = 28)
p63 positive (n = 47)
Total (n = 75)
P-value
62.0 (14.9)
60.2 (16.1)
60.9 (15.6)
0.69
19 9
23 24
42 33
0.19
9 19
24 23
33 42
0.15
Numbers are number (percentage) or median (interquartile range) as appropriate. P-values are odds comparing p63 negative with p63 positive. aSkin, lung, parotid, and brain.
JEM
S3
Table S4. Mutational status of p53, BRAF, and NRAS in melanoma cell lines
Table S5. Culture media and supplements for the melanocytes and established melanoma cell lines
Cell line
Cell line
TP53 status
RGP cells WM35 WT WM1575 WT WM1552C R248Q/+ SBCl2 WT VGP cells WM793 WT WM115 WT WM278 WT VMM39 WT LM3 WT LM4 Y236H/++ LM5 WT Mel 224 WT Mel 505 WT Metastatic melanoma cells LM32 WT LM34 WT LM37 WT Mel 501 WT SK-MEL 5 WT DX3 P278R/+ DX3LT5.1 P278R/+ WM1158 WT WM9 WT WM852 S241P WM983B mt A375M WT C8161 WT SK-MEL 31 WT MALME-3M WT SK-MEL 24 WT HBL WT WM239A WT MM-AN Null UISO-mel-6 Null VGP, vertical growth phase.
BRAF status
NRAS status
V600E V600E V600E WT
WT WT WT Q61K
V600E V600D V600E WT WT V600E V600E WT WT
WT WT WT WT Q61R WT WT Q61R WT
WT V600E WT V600E WT WT WT V600E V600E WT V600E V600E WT WT V600E V600E WT V600D V600E WT
WT WT WT WT WT Q61R Q61R WT WT Q61R WT WT WT WT WT WT WT WT WT Q61R
Culture media and supplements
Human primary melanocytes NHEM, HEMa 254 medium + HMGS-2 (Invitrogen) RGP cells WM35 RPMI + 10% FCS + 1% Gln WM1575 Tu 2% + 10% FCS WM1552C Tu 2% + 10% FCS SBCl2 RPMI + 10% FCS + 1% Gln VGP cells WM793 RPMI + 10% FCS + 1% Gln WM115 RPMI + 10% FCS + 1% Gln WM278 RPMI + 10% FCS + 1% Gln VMM39 RPMI + 10% FCS + 1% Gln LM3 RPMI + 10% FCS + 1% Gln LM4 RPMI + 10% FCS + 1% Gln LM5 RPMI + 10% FCS + 1% Gln Mel 224 RPMI + 10% FCS + 0.5% NEAA + 1% Gln Mel 505 RPMI + 10% FCS + 0.5% NEAA + 1% Gln Metastatic melanoma cells LM32 RPMI + 10% FCS + 1% Gln LM34 RPMI + 10% FCS + 1% Gln LM37 RPMI + 10% FCS + 1% Gln Mel 501 F10 + 10% FCS + 1% Gln SK- MEL 5 MEM + 10% FCS + 1% Gln DX3 RPMI + 10% FCS + 1% Gln DX3-LT5.1 RPMI + 10% FCS + 1% Gln WM1158 RPMI + 10% FCS + 1% Gln WM9 RPMI + 10% FCS + 1% Gln WM852 RPMI + 10% FCS + 1% Gln WM983B RPMI + 10% FCS + 1% Gln A375M RPMI + 10% FCS + 1% Gln C8161 RPMI + 10% FCS+ 20 mM Hepes + 0.1% GS SK-MEL 31 MEM + 10% FCS + 1% Gln MALME-3M RPMI+ 10% FCS + 1% Gln SK-MEL 24 MEM + 10% FCS + 1% Gln HBL MEM + 10% FCS + 1 mM SP + 1% Gln WM239A RPMI + 10% FCS + 1% Gln MM-AN DMEM + 10% FCS + 1% Gln UISO-mel-6 MEM + 15% FCS+ 0.5% NEAA + 1% Gln + 0.1% GS HMGS-2, human melanocytes growth supplement-2; Gln, glutamine; GS, gentamicin sulphate; NEAA, nonessential amino acids; SP, sodium pyruvate; VGP, vertical growth phase.
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Oncogenic p63 is a p53 repressor in melanoma | Matin et al.
