Saussurea tridactyla Sch. Bip.-derived polysaccharides and flavones reduce oxidative damage in ultraviolet B-irradiated HaCaT cells via a p38MAPK-independent mechanism

Figures & data

Table 1 Antibodies and conditions used in Western blotting

Protein	Primary antibody	Primary dilution	MW (kD)	Secondary antibody	Secondary dilution
p-38	Abcam, ab38238	1:800	38	Antirabbit, Zhong Shan Jinqiao Biological Technology Co., Ltd., Beijing, People’s Republic of China	1:5,000
p-53	Abcam, ab194734	1:1,000	53	Antirabbit, Zhong Shan Jinqiao Biological Technology	1:5,000
Bcl-2	Abcam, ab131457	1:1,000	26	Antirabbit, Zhong Shan Jinqiao Biological Technology	1:5,000
Bax	Abcam, ab7977	1:1,000	26	Antirabbit, Zhong Shan Jinqiao Biological Technology	1:5,000
Active-caspase-3	Abcam, ab32042	1:1,000	17	Antirabbit, Zhong Shan Jinqiao Biological Technology	1:5,000
β-actin	Abcam, ab8227	1:1,000	43	Antirabbit, Zhong Shan Jinqiao Biological Technology	1:5,000

Figure 1 Cell viability assay of Saussurea tridactyla Sch. Bip.-derived polysaccharides.

Notes: (A) Cell viability assay of Saussurea tridactyla Sch. Bip.-derived polysaccharides in L-UVB group. (B) Cell viability assay of Saussurea tridactyla Sch. Bip.-derived polysaccharides in H-UVB group. ^#P<0.05, UVB group vs control group. *P<0.05, 10 mg/mL group vs UVB group, 40 mg/mL group vs UVB group. **P<0.05, 40 mg/mL group vs 10 mg/mL group.
Abbreviations: L-UVB, low radiation UVB; H-UVB, high radiation UVB; UVB, ultraviolet B; UVB group, UVB radiation group.

Figure 2 Cell viability assay with Saussurea tridactyla Sch. Bip.-derived flavones in (A) L-UVB group and (B) H-UVB group.

Notes: ^#P<0.05, UVB group vs control group. *P<0.05, 2 mg/mL group vs UVB group, 6 mg/mL group vs UVB group. **P<0.05, 6 mg/mL group vs 2 mg/mL group.
Abbreviations: L-UVB, low radiation UVB; H-UVB, high radiation UVB; UVB, ultraviolet B; UVB group, UVB radiation group.

Figure 3 Trypan blue staining of Saussurea tridactyla Sch. Bip.-derived polysaccharides.

Abbreviations: L-UVB, low radiation UVB; H-UVB, high radiation UVB; UVB, ultraviolet B.

Figure 4 Trypan blue staining of Saussurea tridactyla Sch. Bip.-derived flavones.

Abbreviations: L-UVB, low radiation UVB; H-UVB, high radiation UVB; UVB, ultraviolet B.

Figure 5 SOD activity (A), GSH content (B), MDA content (C), and CAT activity (D) of Saussurea tridactyla Sch. Bip.-derived polysaccharides.

Notes: (A1, B1, C1, D1) show the results of SOD activity, GSH content, MDA content and CAT activity in Saussurea tridactyla Sch. Bip.-derived polysaccharides in L-UVB group. (A2, B2, C2, D2) show the results of SOD activity, GSH content, MDA content and CAT activity in Saussurea tridactyla Sch. Bip.-derived polysaccharides in H-UVB group. ^#P<0.05, UVB group vs control group. *P<0.05, 10 mg/mL group vs UVB group, 40 mg/mL group vs UVB group. **P<0.05, 40 mg/mL group vs 10 mg/mL group.
Abbreviations: L-UVB, low radiation UVB; H-UVB, high radiation UVB; UVB group, UVB radiation group; UVB, ultraviolet B; SOD, superoxide dismutase; GSH, glutathione; MDA, malondialdehyde; CAT, catalase.

Figure 6 SOD activity (A), GSH content (B), MDA content (C), and CAT activity (D) of Saussurea tridactyla Sch. Bip.-derived flavones.

Notes: (A1, B1, C1, D1) show the results of SOD activity, GSH content, MDA content and CAT activity in Saussurea tridactyla Sch. Bip.-derived flavones in L-UVB group. (A2, B2, C2, D2) show the results of SOD activity, GSH content, MDA content and CAT activity in Saussurea tridactyla Sch. Bip.-derived flavones in H-UVB group. ^#P<0.05, UVB group vs control group. *P<0.05, 2 mg/mL group vs UVB group, 6 mg/mL group vs UVB group. **P<0.05, 6 mg/mL group vs 2 mg/mL group.
Abbreviations: L-UVB, low radiation UVB; H-UVB, high radiation UVB; UVB, ultraviolet B; UVB group, UVB radiation group; SOD, superoxide dismutase; GSH, glutathione; MDA, malondialdehyde; CAT, catalase.

Figure 7 Apoptosis assays (A), intracellular Ca²⁺ (B), and mitochondrial membrane potential (Δψ) (C) indicated by flow cytometry of Saussurea tridactyla Sch. Bip.-derived polysaccharides.

Abbreviations: L-UVB, low radiation UVB; H-UVB, high radiation UVB; UVB, ultraviolet B.

Figure 8 Apoptosis assays (A), intracellular Ca²⁺ (B), and mitochondrial membrane potential (Δψ) (C) indicated by flow cytometry of Saussurea tridactyla Sch. Bip.-derived flavones.

Abbreviation: UVB, ultraviolet B.

Figure 9 Expressions of p-38, p-53, Bcl-2, Bax, and active-caspase-3 indicated by Western blotting of Saussurea tridactyla Sch. Bip.-derived polysaccharides.

Notes: ^#P<0.05, UVB group vs control group. *P<0.05, 10 mg/mL group vs UVB group, 40 mg/mL group vs UVB group. **P<0.05, 40 mg/mL group vs 10 mg/mL group.
Abbreviations: L-UVB, low radiation UVB; H-UVB, high radiation UVB; UVB, ultraviolet B; UVB group, UVB radiation group.

Figure 10 Expressions of p-38, p-53, Bcl-2, Bax, and active-caspase-3 indicated by Western blotting of Saussurea tridactyla Sch. Bip.-derived flavones.

Notes: ^#P<0.05, UVB group vs control group. *P<0.05, 6 mg/mL group vs UVB group.
Abbreviations: UVB, ultraviolet B; UVB group, UVB radiation group.

Figure 11 The expressions of p-38, p-53, Bcl-2, Bax, and active-caspase-3 indicated by immunofluorescence staining of Saussurea tridactyla Sch. Bip.-derived polysaccharides.

Abbreviations: L-UVB, low radiation UVB; H-UVB, high radiation UVB; UVB, ultraviolet B.

Figure 12 The expressions of p-38, p-53, Bcl-2, Bax, and active-caspase-3 indicated by immunofluorescence staining of Saussurea tridactyla Sch. Bip.-derived flavones.

Abbreviation: UVB, ultraviolet B.