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. 2024 Dec 20;14(12):632. doi: 10.3390/bios14120632

Table 2.

Changes in the expression levels of candidate marker genes by RNA-Seq analysis and real-time PCR.

Chemical Murine LLNA Category h-CLAT Judgment GAPDH HMOX1 JUN PPP1R15A BTG2 DDIT3 EGR1
NGS NGS PCR Judgment NGS PCR Judgment NGS PCR Judgment NGS PCR Judgment NGS PCR Judgment NGS PCR Judgment
2,4-Dinitrochlorobenzene Extreme p 1.2 5.2 3.2 7.7 9.3 9.3 8.0 3.3 30.7 1.6 1.4 5.4 8.3
3.2 Match 10.0 Match 21.1 Match 4.3 Match 0.7 Match 8.1 Match
2.9 10.0 5.9 2.8 1.4 8.3
1,4-Phenylendiamine Strong p 0.9 83.7 66.3 27.2 19.2 25.6 12.1 10.1 15.7 6.1 5.1 5.1 14.9
69.6 Match 22.8 Match 19.2 Match 16.7 Match 5.5 Match 16.4 Mismatch
61.0 19.2 38.9 28.8 9.6 15.3
Nickel sulfate Moderate p 1.1 3.4 2.3 4.5 2.8 3.1 2.2 4.6 3.9 2.4 1.5 1.1 1.3
2.3 Match 3.0 Match 2.0 Match 6.8 Match 2.2 Match 1.1 Match
2.6 3.1 4.8 5.4 3.9 1.0
2-Mercaptobenzothiazole Moderate p 1.1 63.7 39.7 25.1 27.6 12.3 10.1 3.8 9.3 27.2 33.4 3.2 3.9
37.3 Match 24.9 Match 27.3 Match 6.1 Match 11.6 Mismatch 3.6 Match
22.5 25.4 21.7 5.2 63.1 3.4
R(+)-Limonene Weak p 1.1 61.5 62.2 28.1 54.2 9.1 10.1 3.5 11.1 8.6 9.8 18.6 50.9
46.9 Match 54.9 Match 29.0 Match 4.6 Match 7.4 Match 36.5 Match
54.6 63.6 17.8 6.1 18.8 32.9
Imidazolidinyl urea Weak p 1.2 7.4 3.9 80.0 65.3 15.6 10.9 13.5 15.8 2.5 2.2 8.3 10.9
4.1 Match 46.8 Match 13.2 Match 18.6 Match 4.6 Match 8.3 Match
4.2 51.9 17.9 24.1 3.3 8.6
Isopropanol non-sensitizer n 1.0 1.3 1.6 1.4 0.8 1.2 1.1 1.4 1.6 1.7 2.3 0.6 0.8
1.5 Match 0.7 Match 0.9 Match 1.5 Match 3.3 Match 0.6 Match
1.5 0.9 1.8 1.6 3.7 0.5
Glycerol non-sensitizer n 1.0 1.5 1.4 1.2 0.8 1.0 1.0 0.9 1.4 1.1 1.1 1.1 1.6
1.6 Match 1.2 Match 1.0 Match 1.3 Match 1.2 Match 1.0 Match
0.9 0.8 1.1 0.8 1.6 1.0
4-Aminobenzoic acid non-sensitizer n 1.1 1.3 0.8 0.6 0.8 1.0 0.8 1.4 1.7 0.9 0.8 0.5 0.8
0.8 Match 0.6 Match 2.0 Match 1.1 Match 0.3 Match 0.5 Match
0.9 0.8 0.9 4.5 1.2 0.6
Chemical Murine LLNA Category h-CLAT Judgment GAPDH GADD45B PMAIP1 SAT1 ULBP2 CCR2 ICMT
NGS NGS PCR Judgment NGS PCR Judgment NGS PCR Judgment NGS PCR Judgment NGS PCR Judgment NGS PCR Judgment
2,4-Dinitrochlorobenzene Extreme p 1.2 7.3 8.1 7.7 6.9 2.4 0.5 7.1 1.6 0.1 0.2 0.5 0.5
7.4 Match 6.5 Match 16.6 Mismatch 8.5 Match 0.3 Mismatch 0.5 Match
5.8 9.3 1.7 4.6 0.2 0.7
1,4-Phenylendiamine Strong p 0.9 14.6 18.3 5.6 4.3 16.5 8.4 8.9 3.7 0.0 0.1 0.3 0.3
12.1 Match 2.9 Match 14.4 Match 1.6 Mismatch 0.0 Match 0.5 Match
25.6 9.9 24.1 12.6 0.0 0.4
Nickel sulfate Moderate p 1.1 0.9 1.0 3.6 2.8 2.3 1.5 1.4 0.5 0.6 0.6 0.6 0.6
0.7 Match 2.6 Match 2.0 Match 0.3 Mismatch 0.5 Match 0.8 Match
1.4 4.3 3.5 1.8 0.7 0.8
2-Mercaptobenzothiazole Moderate p 1.1 6.5 7.9 4.5 4.9 12.7 2.2 4.2 0.9 0.1 0.1 0.4 0.6
7.2 Match 3.9 Match 27.3 Mismatch 4.6 Mismatch 0.1 Match 0.6 Match
8.6 6.5 27.7 8.6 0.2 0.6
R(+)-Limonene Weak p 1.1 3.4 3.5 3.9 3.9 8.0 3.6 7.5 2.2 0.1 0.5 0.2 0.5
3.5 Match 4.2 Match 44.6 Mismatch 11.6 Mismatch 0.3 Mismatch 0.9 Mismatch
3.4 8.3 17.9 17.6 0.3 0.7
Imidazolidinyl urea Weak p 1.2 12.0 6.0 19.4 14.5 15.0 12.3 24.3 21.3 0.0 0.0 0.1 0.2
5.9 Mismatch 15.8 Match 18.5 Match 11.8 Match 0.0 Match 0.2 Mismatch
9.3 32.7 13.1 26.7 0.0 0.2
Isopropanol non-sensitizer n 1.0 1.2 1.4 1.2 2.1 1.3 2.1 1.1 1.6 0.8 1.0 1.0 0.9
0.7 Match 2.2 Match 3.7 Mismatch 0.8 Match 0.7 Match 1.2 Match
1.9 2.1 2.8 3.0 1.2 1.1
Glycerol non-sensitizer n 1.0 0.9 1.1 1.1 1.3 1.0 1.0 0.8 0.9 1.2 1.8 0.9 0.7
0.7 Match 1.0 Match 1.5 Match 0.4 Match 1.1 Match 1.5 Match
1.1 2.0 1.9 2.1 1.1 1.1
4-Aminobenzoic acid non-sensitizer n 1.1 1.1 1.0 1.3 0.9 1.1 0.3 1.6 0.4 0.7 0.6 0.9 3.3
0.7 Match 0.8 Match 3.8 Match 1.2 Match 0.8 Match 0.9 Match
0.6 8.2 1.8 2.1 0.5 1.0

The potency of the sensitizers in the murine LLNA [3,19] is classified as extreme, strong, moderate or weak. In the judgment by h-CLAT [7,26], if either CD86 or CD54 was positive, the test chemical is also judged as positive, “p”. Otherwise, it is indicated by “n”. If the results of new generation sequencing (NGS) and real-time PCR are consistent, it is marked as “Match”; if not, it is marked as “Mismatch”.