Hsp90 Inhibitors in FF Cells of Control of Control of Control 50 Vemurafenib (𝜇M Vemurafenib (𝜇M Vemurafenib (𝜇M of Control of Control of Control 0 0.125 0

-8 -6 -4 0

50 100

Log₁₀ (Hsp90i) [M]

Cell Viability (%)

KU758 XL888 WGA-TA

IC50 (µM)

XL888 113.3

KU758 4.05

WGA-TA 2.28

Supplemental Figure 1 –Dose-response curves of each Hsp90i in FF cells are shown with corresponding 24 hrs IC₅₀ values tabulated in micromolar concentrations.

Hsp90 Inhibitors in FF Cells

50 25

12.5

0 0

50 100

% of Control

0 50 100

% of Control

0 50 100

% of Control

50 Vemurafenib (𝜇M)

25

12.5 0

50 Vemurafenib (𝜇M)

25

12.5 0

Vemurafenib (𝜇M)

0 50 100

% of Control

0 50 100

% of Control

0 50 100

% of Control

0 0.125

0. 25 0. 5

KU758 ( 𝜇M)

0 0.125

0. 25 0. 5

KU758 ( 𝜇M)

0 0.025

0.05 0.1

WGA-TA (𝜇M)

0 0.025

0.05 0.1

WGA-TA (𝜇M)

0 0.075

0.15 0.3

XL888 ( 𝜇M)

0 0.075

0.15 0.3

XL888 ( 𝜇M)

0.5 0.25

0.125 0

Cobimetinib (𝜇M)

0.5 0.25

0.125 0

Cobimetinib (𝜇M)

0.5 0.25

0.125 0

Cobimetinib (𝜇M)

0 0.125

0. 25

0. 5 50

Vemurafenib (𝜇M)

25

12.5 0

Synergy Antagonism

0 50 100

% of Control

Cobimetinib (𝜇M)

Supplemental Figure 2 – The combination effect of Hsp90 inhibitor combinations in UACC-62. Ve+Cb combination (FDA approved) is shown for comparison.

Supplemental Figure 3 - Dose-response curves of Hsp90i KU758 in melanoma cells are shown with corresponding 24 hrs IC50 values tabulated in micromolar concentrations.

Skmel103 Skmel173 Skme119

I

•

�

100 100

I

?;, 75 'l, ⁷⁵

·� 5

⁵⁰ '/.

,,

₅₀

� ^;!.

25 25

0 0

-7.0 -6.5 -6.0 -5.5 -5.0 -7,0 -6.5 -6.0 -5.5 -5.0

-7,0 -6.5 -6.0 -5.5 -5.0

I.og[KU7:'8) M L,g[K.l.i75S] M I.og[KU758] M

Supplemental Figure 4 – Scratch and Boyden chamber assays in UACC-62. A) KU758 inhibitor combinations decreased cell migration (red lines indicate scratched cell-free area). B) quantification of the percent of cell migration at 24 hours after Hsp90i and MAPKi combinations. p values are recorded as significance between t = 0 and 24 hrs. C) The number of migrated cells after inhibitor single-agent and combination treatments.

0%

20%

40%

60%

80%

100%

Untr. Ve Cb Ve+Cb XL888 XL888+Ve XL888+Cb WGA-TA WGA-TA+Ve WGA-TA+Cb KU758 KU758+Ve KU758+Cb

% Migrated

0 100 200 300 400 500

Untr. Ve Cb Ve+Cb XL888 XL888+Ve XL888+Cb WGA-TA WGA-TA+Ve WGA-TA+Cb KU758 KU758+Ve KU758+Cb

Migrated Cell (#)

B) A)

0 hrs24 hrs

Untreated Cb Ve Ve + Cb KU758 KU758 + Cb KU758 + Ve

C)

* p < 0.05

** p < 0.001

*

* *

*

* * *

*

*

* p < 0.05

** p < 0.001

**

* *

**

** *